GIP agonist compounds and methods

ABSTRACT

The present invention relates to acylated GIP analogs which have GIP agonist activity, and their use in the treatment of metabolic disorders.

FIELD OF THE INVENTION

The invention relates to compounds having agonist activity at the GIPreceptor, and to their use in the treatment of metabolic disorders.

BACKGROUND OF THE INVENTION

Diabetes and obesity are increasing health problems globally and areassociated with various other diseases, particularly cardiovasculardiseases (CVD), obstructive sleep apnea, stroke, peripheral arterydisease, microvascular complications and osteoarthritis. There are 246million people worldwide with diabetes, and by 2025 it is estimated that380 million will have diabetes. Many have additional cardiovascular riskfactors including high/aberrant LDL and triglycerides and low HDL.Cardiovascular diseases account for about 50% of the mortality in peoplewith diabetes, and the morbidity and mortality rates relating to obesityand diabetes underscore the medical need for efficacious treatmentoptions.

Glucose-dependent insulinotropic polypeptide (“GIP”, also known as“gastric inhibitory polypeptide”) is a 42-residue peptide secreted byenteroendocrine K-cells of the small intestine into the bloodstream inresponse to oral nutrient ingestion. GIP inhibits the secretion ofgastric acid, and it has been shown to be a potent stimulant for thesecretion of insulin from pancreatic beta cells after oral glucoseingestion (the “incretin effect”) (Creutzfeldt, W., et al, 1979,Diabetologia, 16:75-85).

Insulin release induced by the ingestion of glucose and other nutrientsis due to both hormonal and neural factors (Creutzfeldt, W., et al,1985, Diabetologia, 28:565-573). Several gastrointestinal regulatorypeptides have been proposed as incretins, and among these candidates,only GIP and glucagon-like peptide 1 (“GLP-1”) appear to fulfill therequirements to be considered physiological stimulants of postprandialinsulin release (Nauck, et al, 1989, J. Clin. Endocrinol Metab.,69:654-662). It has been shown that the combined effects of GIP andGLP-1 are sufficient to explain the full incretin effect of theenteroinsular axis (Fehmann, H. C, et al, 1989, FEBS Lett, 252:109-112).

As is well known to those skilled in the art, the known and potentialuses of GIP are varied and multitudinous. Thus, the administration ofthe compounds of this invention for purposes of eliciting an agonisteffect can have the same effects and uses as GIP itself. These varieduses of GIP may be summarized as follows: treating a disease selectedfrom the group consisting of type 1 diabetes, type 2 diabetes (Visboll,T., 2004, Dan. Med. Bull, 51:364-70), insulin resistance (WO2005/082928), obesity (Green, B. D., et al, 2004, Current PharmaceuticalDesign, 10:3651-3662), metabolic disorder (Gault, V. A., et al, 2003,Biochem. Biophys. Res. Commun., 308:207-213), central nervous systemdisease, neurodegenerative disease, congestive heart failure,hypoglycemia, and disorders wherein the reduction of food intake andweight loss are desired. In pancreatic islets, GIP not only enhancesinsulin secretion acutely, but it also stimulates insulin productionthrough enhancement of proinsulin transcription and translation (Wang,et al, 1996, Mol Cell. Endocrinol, 116:81-87) and enhances the growthand survival of pancreatic beta cells (Trumper, et al, 2003, Diabetes,52:741-750). In addition to effects on the pancreas to enhance insulinsecretion, GIP also has effects on insulin target tissues directly tolower plasma glucose: enhancement of glucose uptake in adipose (Eckel,et al, 1979, Diabetes, 28: 1141-1142) and muscle (O'Harte, et al, 1998,J. Endocrinol, 156:237-243), and inhibition of hepatic glucoseproduction (Elahi, D., et al, 1986, Can. J. Physiol. Pharmacol, 65:A18).

Recently, it has been reported that body weight loss associated withGLP-1 agonist treatment, is enhanced when GLP-1 and GIP areco-administered (Finan, Sci Transl Med. 2013; 5(209):209ra151. Irwin Net al, 2009, Regul Pept; 153: 70-76. Gault et al, 2011, Clin Sci (Lond);121:107-117). For instance, Finan and colleges demonstrated significantbody weight loss in diet-induced obese (DIO) mice after sub-chronicco-administration with an acylated GIP agonist and an acylated GLP-1agonist. The co-administration decreased body weight and fat mass to agreater extent than either mono-agonist alone. Evidence also suggeststhat GLP-1 and GIP have additive effects on glycemic control (Gault etal, 2011, Clin Sci (Lond); 121:107-117). A study by Gault et al showedthat sub-chronic co-administration with a GLP-1 analogue, and anacylated GIP analogue resulted in greater glucose-lowering andinsulinotropic actions during an intraperitoneal glucose tolerance testin ob/ob mice than injection with the GLP-1 agonist or the GIP agonistalone. Thus, GIP agonists may be particular effective in improvingglycemic control and reducing body weight when they are administered incombination with a GLP-1 receptor agonist (as part of the samepharmaceutical formulation or as separate formulations).

The use of unmodified GIP as a therapeutic, however, is limited by theshort in vivo half-life of about 2 minutes (Said and Mutt, 1970,Science, 169:1217-1218). In serum, both incretins, GIP and GLP-1, aredegraded by dipeptidyl peptidase IV (“DPPIV”). Improving the stabilityof GIP to proteolysis not only maintains the activity of GIP at itsreceptor but, more importantly, prevents the production of GIPfragments, some of which act as GIP receptor antagonists (Gault, et al.,2002, J. Endocrinol, 175:525-533). Reported modifications have includedprotection of the N-terminus of GIP from proteolysis by DPPIV throughmodification of the N-terminal tyrosine (O'Harte, et al, 2002,Diabetologia, 45: 1281-1291), mutation of the alanine at position 2(Hinke, et al, 2002, Diabetes, 51:656-661), mutation of glutamic acid atposition 3 (Gault, et al, 2003, Biochem. Biophys. Res. Commun.,308:207-213), and mutation of alanine at position 13 (Gault, et al,2003, Cell Biol. International, 27:41-46), The following patentapplications have been filed related to the effects of GIP analogues onthe function of various target organs and their potential use astherapeutic agents: PCT publication WO 00/58360 discloses peptidylanalogues of GIP which stimulate the release of insulin. In particular,this application discloses specific peptidyl analogues comprising atleast 15 amino acid residues from the N-terminal end of GIP(I-42. PCTpublication WO 03/082898 discloses C-terminal truncated fragments andN-terminal modified analogues of GIP, as well as various GIP analogueswith a reduced peptide bond or alterations of the amino acids close tothe DPPFV-specific cleavage site. This application further disclosesanalogues with different linkers between potential receptor bindingsites of GIP. The compounds of this application are alleged to be usefulin treating GIP-receptor mediated conditions, such as non-insulindependent diabetes mellitus and obesity. Moreover, among othertherapeutic effects of the compounds of the present invention asillustrated herein, tighter control of plasma glucose levels may preventlong-term diabetic complications, thereby providing an improved qualityof life for patients. In addition to improving blood glucose control,GIP may also enhance GLP-1-mediated body weight loss.

Conjugation of GIP analogues to e.g, PEG(poly ethylene glycol) has beenshown to extent in vivo half-life, but potential side-effects ofpegylated pharmaceutical products such as inteferon-beta and ribavirinhas been reported (J Clin Gastroenterol. 2004 September; 38(8):717-22,Gut 2006; 55:1350-1359 doi:10.1136/qut.2005.076646).

Thus, there still exists a need for improved and safe analogues of GIP,which are stable in formulation and have long in vivo half-life,resulting from decreased susceptibility to proteolysis and decreasedclearance, while maintaining binding affinity to a GIP receptor toelicit agonistic effects.

SUMMARY OF THE INVENTION

The present invention concerns GIP analogues which may have the propertyof an altered GIP activity, as assessed in in vitro efficacy assays andan altered, preferably increased terminal elimination half-life (T½), asassessed in in vivo studies in mice.

It has been found that GIP receptor agonists of the present inventionare superior to existing GIP analogues because the GIP agonists offerlong terminal half-lifes. The GIP analogues may thus be used astherapeutics for metabolic disorders including, but not limited to, type2 diabetes mellitus, obesity and related disorders.

The invention provides in a first aspect a GIP analogue represented bythe general Formula I:R¹-Tyr-X2-Glu-Gly-Thr-Phe-Ile-Ser-Asp-X10-X11-X12-Glu-Leu-X15-X16-X17-X18-X19-X20-X21-Phe-X23-X24-X25-Leu-X27-X28-X29-Y1-Y2-R²  (I)(SEQ ID NOs: 42 & 86)whereinR¹ is H—, Ac or pGlu pyroglutamic acid (pGlu;(S)-(−)-2-pyrrolidone-5-carboxylic acid), C₁₋₄ alkyl, acetyl, formyl,benzoyl and trifluoroacetyl,X2 is Aib, Ala, D-Ala, Gly, Ser, N-Me-Ser, Ac3c, Ac4c or Ac5c;X10 is Tyr, Leu or Ser;X11 is Ser or Leu;X12 is Lys, Ψ or Ile;X15 is Asp or Glu;X16 is Ser, Glu, Lys or Ψ;X17 is Ile, Lys, Gln, Arg or Ψ;X18 is His, Arg or Ala;X19 is Gln, Lys, Ala or Glu;X20 is Gln, Lys, Ala, His or Arg;X21 is Ala, Leu, Asp or Glu;X23 is Val or Ile;X24 is Asn or Glu;X25 is Tyr or Trp;X27 is Leu, Glu, Ser, Lys or Val;X28 is Ala, Ser or Arg;X29 is Aib, Gly, Ala, Gln, Thr, Ser or Lys or is absent;Y1 is Lys-Gly, Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser (SEQ ID NO: 43),Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 44),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser (SEQ ID NO: 45),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 46),Gly-Lys-Lys-Asn-Asp-Trp-Lys-His-Asn-Ile-Thr-Gln (SEQ ID NO: 47) orabsent;Y2 is Ψ or is absent;R² is —NH₂ or —OH;wherein Ψ is a residue independently selected from Lys, Arg, Orn and Cysand wherein the side chain of said residue is conjugated to a lipophilicsubstituent;and wherein the GIP analogue contains one and only one residue Ψ;or a pharmaceutically acceptable salt or solvate thereof.

In one aspect, R¹ is H—, Ac or pGlu.

Combinations of residues which may be present at some of the variablepositions of Formula I include:

Aib2, Asp15, Lys20;

Aib2, Asp15, Arg20;

Aib2, Asp15, Arg20, Ile23;

Aib2, Ile12, Asp15, Arg20, Ile23, Glu24;

Ile12, Asp15, Ile23;

Ile12, Asp15, Ile23, Glu24;

Ile12, Asp15, Ala21, Ile23;

Aib2, Ala21, Ile23, Glu24;

Aib2, Asp15, Ile23;

Aib2, Asp15, Arg20, Ile23, Gln29;

Aib2, Asp15, Arg20, Gly29;

Aib2, Asp15, Ile17, Arg20, Gly29;

Aib2, Asp15, Ile17, Lys20, Gly29;

DAla2, Asp15, Ile23;

DAla2, Asp15, Ile23, Ala28;

Aib2, Asp15, Ile17, Lys20, Ala28;

Asp15, Ile23, Glu24;

N-Me-Ser2, Asp15, Lys20;

N-Me-Ser2, Asp15, Arg20;

N-Me-Ser2, Asp15, Arg20, Ile23;

N-Me-Ser2, Ile12, Asp15, Arg20, Ile23, Glu24;

N-Me-Ser2, Ala21, Ile23, Glu24;

N-Me-Ser2, Asp15, Ile23;

N-Me-Ser2, Asp15, Arg20, Ile23, Gln29;

N-Me-Ser2, Asp15, Arg20, Gly29;

N-Me-Ser2, Asp15, Ile17, Arg20, Gly29;

N-Me-Ser2, Asp15, Ile17, Lys20, Gly29;

N-Me-Ser2, Asp15, Ile23;

N-Me-Ser2, Asp15, Ile23, Ala28;

Ac3c2, Asp15, Lys20;

Ac3c2, Asp15, Arg20;

Ac3c2, Asp15, Arg20, Ile23;

Ac3c2, Ile12, Asp15, Arg20, Ile23, Glu24;

Ac3c2, Ala21, Ile23, Glu24;

Ac3c2, Asp15, Ile23;

Ac3c2, Asp15, Arg20, Ile23, Gln29;

Ac3c2, Asp15, Arg20, Gly29;

Ac3c2, Asp15, Ile17, Arg20, Gly29;

Ac3c2, Asp15, Ile17, Lys20, Gly29;

Ac3c2, Asp15, Ile23;

Ac3c2, Asp15, Ile23, Ala28;

Ac4c2, Asp15, Lys20;

Ac4c2, Asp15, Arg20;

Ac4c2, Asp15, Arg20, Ile23;

Ac4c2, Ile12, Asp15, Arg20, Ile23, Glu24;

Ac4c2, Ala21, Ile23, Glu24;

Ac4c2, Asp15, Ile23;

Ac4c2, Asp15, Arg20, Ile23, Gln29;

Ac4c2, Asp15, Arg20, Gly29;

Ac4c2, Asp15, Ile17, Arg20, Gly29;

Ac4c2, Asp15, Ile17, Lys20, Gly29;

Ac4c2, Asp15, Ile23;

Ac4c2, Asp15, Ile23, Ala28;

Ac5c2, Asp15, Lys20;

Ac5c2, Asp15, Arg20;

Ac5c2, Asp15, Arg20, Ile23;

Ac5c2, Ile12, Asp15, Arg20, Ile23, Glu24;

Ac5c2, Ala21, Ile23, Glu24;

Ac5c2, Asp15, Ile23;

Ac5c2, Asp15, Arg20, Ile23, Gln29;

Ac5c2, Asp15, Arg20, Gly29;

Ac5c2, Asp15, Ile17, Arg20, Gly29;

Ac5c2, Asp15, Ile17, Lys20, Gly29;

Ac5c2, Asp15, Ile23; or

Ac5c2, Asp15, Ile23, Ala28.

The invention provides in a further aspect a GIP analogue represented bythe general Formula II:R¹-Tyr-X2-Glu-Gly-Thr-Phelle-Ser-Asp-Tyr-Ser-X12-Glu-Leu-X15-X16-X17-X18-X19-X20-X21-Phe-X23-X24-X25-Leu-X27-X28-X29-Y1-Y2-R²  (II)(SEQ ID NOs: 48 & 87)whereinR¹ is H—, Ac or pGlu;X2 is Aib, Ala, D-Ala or Gly;X12 is Lys, Ψ or Ile;X15 is Asp or Glu;X16 is Ser, Glu, Lys or Ψ;X17 is Ile, Lys, Gln, Arg or Ψ;X18 is His, Arg or Ala;X19 is Gln or Ala;X20 is Gln, Lys, Ala, His or Arg;X21 is Ala, Asp or Glu;X23 is Ile or Val;X24 is Asn or Glu;X25 is Tyr or Trp;X27 is Leu, Glu, Ser, Lys or Val;X28 is Ala, Ser or Arg;X29 is Aib, Gly, Ala, Gln, Thr, Ser or Lys or is absent;Y1 is Lys-Gly, Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro (SEQ ID NO: 49),Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 44),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser (SEQ ID NO:45),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 46),Gly-Lys-Lys-Asn-Asp-Trp-Lys-His-Asn-Ile-Thr-Gln (SEQ ID NO: 47) orabsent;Y2 is Ψ or is absent;R² is —NH₂ or —OH;wherein Ψ is a Lys residue wherein the side chain of said Lys residue isconjugated to a lipophilic substituent;and wherein the GIP analogue contains one and only one residue ip;or a pharmaceutically acceptable salt or solvate thereof.

Combinations of residues which may be present at some of the variablepositions of Formula II include:

Aib2, Lys12, Asp15, Lys20;

Aib2, Lys12, Asp15, Arg20;

Aib2, Asp15, Arg20;

Aib2, Ile12, Asp15, Arg20, Glu24;

Ile12, Asp15, Ile23;

Ile12, Asp15, Glu24;

Ile12, Asp15, Ala21;

Aib2, Lys12, Ala21, Glu24;

Aib2, Lys12, Asp15;

Aib2, Lys12, Asp15, Arg20, Gln29;

Aib2, Lys, 12, Asp15, Arg20, Gly29;

Aib2, Lys12, Asp15, Ile17, Arg20, Gly29;

Aib2, Asp15, Ile17, Lys20, Gly29;

DAla2, Asp15;

DAla2, Asp15, Ala28;

Aib2, Asp15, Ile17, Lys20, Ala28;

Asp15, Glu24;

Ala2, Lys12, Asp15, Lys20;

Ala2, Lys12, Asp15, Arg20;

Ala2, Asp15, Arg20;

Ala2, Ile12, Asp15, Arg20, Glu24;

Ala2, Ile12, Asp15, Ile23;

Ala2, Ile12, Asp15, Glu24;

Ala2, Ile12, Asp15, Ala21;

Ala2, Lys12, Ala21, Glu24;

Ala2, lys12, Asp15;

Ala2, Lys12, Asp15, Arg20, Gln29;

Ala2, Lys12, Asp15, Arg20, Gly29;

Ala2, Lys12, Asp15, Ile17, Arg20, Gly29;

Ala2, Asp15, Ile17, Lys20, Gly29;

Ala2, Asp15;

Ala2, Asp15, Ala28;

Ala2, Asp15, Ile17, Lys20, Ala28;

Gly2, Lys12, Asp15, Lys20;

Gly2, Lys12, Asp15, Arg20;

Gly2, Asp15, Arg20;

Gly2, Ile12, Asp15, Arg20, Glu24;

Gly2, Ile12, Asp15, Ile23;

Gly2, Ile12, Asp15, Glu24;

Gly2, Ile12, Asp15, Ala21;

Gly2, Lys12, Ala21, Glu24;

Gly2, Lys12, Asp15;

Gly2, Lys12, Asp15, Arg20, Gln29;

Gly2, Lys12, Asp15, Arg20, Gly29;

Gly2, Lys12, Asp15, Ile17, Arg20, Gly29;

Gly2, Asp15, Ile17, Lys20, Gly29;

Gly2, Asp15;

Gly2, Asp15, Ala28;

Gly2, Asp15, Ile17, Lys20, Ala28; or

Gly2, Asp15, Glu24.

The invention provides in a further aspect a GIP analogue represented bythe general Formula III:R¹-Tyr-Aib-Glu-Gly-Thr-Phelle-Ser-Asp-Tyr-Ser-Ile-Glu-Leu-X15-X16-X17-X18-X19-X20-X21-Phe-Val-X24-X25-Leu-Leu-Ala-X29-Y1-Y2-R²  (III)(SEQ ID NOs: 50 & 88)whereinR¹ is H—, Ac or pGlu;X15 is Asp or Glu;X16 is Lys or Ψ;X17 is Ile or Ψ;X18 is His or Ala;X19 is Gln or Ala;X20 is Gln, Lys or Arg;X21 is Ala, Asp or Glu;X24 is Asn or Glu;X25 is Tyr or Trp;X28 is Ala, Ser or Arg;X29 is Gln or is absent;Y1 is Lys-Gly, Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser (SEQ ID NO: 43),Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 44),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser (SEQ ID NO: 45),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 46),Gly-Lys-Lys-Asn-Asp-Trp-Lys-His-Asn-Ile-Thr-Gln (SEQ ID NO: 47) orabsent;Y2 is Ψ or is absent;R² is —NH₂ or —OH;wherein Ψ is a residue independently selected from Lys, Arg, Orn and Cysand wherein the side chain of said residue is conjugated to a lipophilicsubstituent;and wherein the GIP analogue contains one and only one residue Ψ;or a pharmaceutically acceptable salt or solvate thereof.

Combinations of residues which may be present at some of the variablepositions of Formula III include:

Asp15, Lys20;

Asp15, Arg20;

Asp15, Arg20, Glu24;

Asp15, Lys 16;

Asp15, Lys 16, Glu24;

Asp15, Ψ16, Ala21;

Ala21,Glu24;

Asp15, Arg20, Gln29;

Asp15, Arg20, Gly29;

Asp15, Ile17, Arg20, Gly29;

Asp15, Ile17, Lys20, Gly29;

Asp15Ala28;

Asp15, Ile17, Lys20, Ala28;

Asp15, Ile23, Glu24;

Asp15, Ψ17, Lys20;

Asp15, Ψ17, Arg20;

Asp15, Ψ17, Arg20

Asp15, Ψ17, Arg20, Glu24;

Asp15, Lys16, Ψ17;

Asp15, Lys16, Ψ17, Glu24;

Asp15, Ψ17, Ala21;

Ala21, Ψ17, Glu24;

Asp15, Asp15, Ψ17, Arg20, Gln29;

Asp15, Ψ17, Arg20, Gly29;

Asp15, Ile17, Arg20, Gly29;

Asp15, Ile17, Lys20, Gly29;

Asp15; Ψ17;

Asp15, Ψ17, Ala28;

Asp15, Ile17, Lys20, Ala28; or

Asp15, Ψ17, Ile23, Glu24.

The GIP-analogue may have the formula R1-Z—R2 where R1 and R2 are asdefined above and Z has the sequence:

(SEQ ID NO: 51) Y-Aib-EGTFISDYSIELDKΨHQQDFVNWLLAQGPSSGAPPPS;(SEQ ID NO: 52) Y-Aib-EGTFISDYSIELDΨIHQQDFVNWLLAQGPSSGAPPPS;(SEQ ID NO: 53) Y-Aib-EGTFISDYSIELEKΨHQQDFVNWLLAQGPSSGAPPPS;(SEQ ID NO: 54) Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPSΨ;(SEQ ID NO: 55) Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQΨ; (SEQ ID NO: 56)Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQKGΨ; (SEQ ID NO: 57)Y-Aib-EGTFISDYSIELDKΨHQQDFVNYLLAQGPSSGAPPPS; (SEQ ID NO: 58)Y-Aib-EGTFISDYSIELDKΨHQQDFVNWLLAQGPSSGAPPPS; (SEQ ID NO: 69)Y-Aib-EGTFISDYSIELDKΨAAQDFVNWLLAQGPSSGAPPPS; (SEQ ID NO: 60)Y-Aib-EGTFISDYSIELEKΨAAKEFVNWLLAQGPSSGAPPPS; (SEQ ID NO: 61)Y-Aib-EGTFISDYSIELEKΨAQRAFVEWLLAQGPSSGAPPPS; (SEQ ID NO: 62)Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQGPSSGAPPPSΨ; (SEQ ID NO: 63)Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQΨ; (SEQ ID NO: 64)Y-Aib-EGTFISDYSIELDKΨAAQDFVNWLLAGPSSGAPPPS; (SEQ ID NO: 65)Y-Aib-EGTFISDYSIELDKIAAQDFVNWLLAGPSSGAPPPSΨ; (SEQ ID NO: 66)Y-Aib-EGTFISDYSIELDKKΨAQRAFVEWLLAQGPSSGAPPPS; (SEQ ID NO: 67)Y-Aib-EGTFISDYSIELDKKΨAQRAFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 68)Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 69)Y-Aib-EGTFISDYSIELDKIAQKEFIEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 70)Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 71)Y-Aib-EGTFISDYSIELDKIAAQDFVEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 72)Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAQGPSSGAPPPSKΨ; (SEQ ID NO: 73)Y-Aib-EGTFISDYSIELDKKΨAAQAFVNWLLAGPSSGAPPPS; (SEQ ID NO: 74)Y-Aib-EGTFISDYSIELDKKΨAAQDFVNWLLAAGPSSGAPPPS, (SEQ ID NO: 75)Y-Aib-EGTFISDYSIELDKKΨAAQDFINWLLAGPSSGAPPPS, (SEQ ID NO: 76)Y-Aib-EGTFISDYSIELDKKΨAAQDFIEWLLAGPSSGAPPPS, (SEQ ID NO: 77)Y-Aib-EGTFISDYSIELDKKΨAAQDFIEWLLAGPSSGAPPPS, (SEQ ID NO: 78)Y-Aib-EGTFISDYSIELDKΨIAQRAFIEWLLAQGPSSGAPPPS, (SEQ ID NO: 79)Y-Aib-EGTFISDYSKΨELDKIAQRAFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 80)Y-DAla-EGTFISDYSIELDKKΨAQRAFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 81)Y-DAla-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 82)Y-Aib-EGTFISDYSIELDKKΨAAQDFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 83)Y-Aib-EGTFISDYSIELDKKΨAAQDFINWLLAQGPSSGAPPPS,  or (SEQ ID NO: 84)Y-Aib-EGTFISDYSIELDKKΨAAQAFIEWLLAQGPSSGAPPPS.

The GIP-analogue may have the formula R1-Z—R2 where R1 and R2 are asdefined above and Z has the sequence

Y-Aib-EGTFISDYSIELDK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 1);Y-Aib-EGTFISDYSIELD-K(Hexadecanoyl-isoGlu)-IHQQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 2);Y-Aib-EGTFISDYSIELEK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 3);Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 4);Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-K(Hexadecanoyl-isoGlu) (SEQ ID NO: 5);Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQ-K(Hexadecanoyl-isoGlu) (SEQ ID NO: 6);Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQKG-K(Hexadecanoyl-isoGlu) (SEQ ID NO: 7);Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNYLLAQGPSSGAPPPS (SEQ ID NO: 8);Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 9);Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 10);Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAKEFVNWLLAQGPSSGAPPPS (SEQ ID NO: 11);Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS (SEQ ID NO: 12);Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 13);Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQ-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 14);H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAGPSSGAPPPS (SEQ ID NO: 15);Y-Aib-EGTFISDYSIELDKIAAQDFVNWLLAGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 16);Y-A1b-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS (SEQ ID NO: 17);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 18);Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 19);Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS (SEQ ID NO: 20);Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS (SEQ ID NO: 21);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS (SEQ ID NO: 22);Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 23);Y-Aib-EGTFISDYSIELDKIAQKEFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 24);Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 25);Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3) (SEQ ID NO: 26);Y-Aib-EGTFISDYSIELDKIAAQDFVEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 27);Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAQGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 28);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyn-isoGlu-Peg3-Peg3)-AAQAFVNWLLAGPSSGAPPPS (SEQ ID NO: 29);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAAGPSSGAPPPS (SEQ ID NO: 30);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAGPSSGAPPPS (SEQ ID NO: 31);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS (SEQ ID NO: 32);Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS (SEQ ID NO: 33);Y-Aib-EGTFISDYSIELD-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-IAQRAFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 34)Y-Aib-EGTFISDYS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-ELDKIAQRAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 35);Y-DAla-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 36);Y-DAla-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3) (SEQ ID NO: 37);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAQGPSSGAPPPS (SEQ ID NO: 38);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAQGPSSGAPPPS (SEQ ID NO: 39);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 40); orY-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 41).

The GIP-analogue may be

(SEQ ID NO: 1) (Compound 1)H-Y-Aib-EGTFISDYSIELDK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS-NH₂;(SEQ ID NO: 2) (Compound 2)H-Y-Aib-EGTFISDYSIELD-K(Hexadecanoyl-isoGlu)-IHQQDFVNWLLAQGPSSGAPPPS-NH₂;(SEQ ID NO: 3) (Compound 3)H-Y-Aib-EGTFISDYSIELEK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS-NH₂;(SEQ ID NO: 4) (Compound 4)H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 5) (Compound 5)H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-K(Hexadecanoyl-isoGlu)-NH₂;(SEQ ID NO: 6) (Compound 6)H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQ-K(Hexadecanoyl-isoGlu)-NH₂;(SEQ ID NO: 7) (Compound 7)H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQKG-K(Hexadecanoyl-isoGlu)-NH₂;(SEQ ID NO: 8) (Compound 8)H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNYLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 9) (Compound 9)H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 10) (Compound 10)H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 11) (Compound 11)H-Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAKEFVNWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 12) (Compound 12)H-Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 13) (Compound 13)H-Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 14) (Compound 14)H-Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQ-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 15) (Compound 15)H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 16) (Compound 16)H-Y-Aib-EGTFISDYSIELDKIAAQDFVNWLLAGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 17) (Compound 17)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 18) (Compound 18)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 19) (Compound 19)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAF1EWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 20) (Compound 20)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 21) (Compound 21)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS-NH₂; (SEQ ID NO: 22) (Compound 22)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS-NH₂; (SEQ ID NO: 23) (Compound 23)H-Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 24) (Compound 24)H-Y-Aib-EGTFISDYSIELDKIAQKEFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 25) (Compound 25)H-Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 26) (Compound 26)H-Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyI)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-NH₂; (SEQ ID NO: 27) (Compound 27)H-Y-Aib-EGTFISDYSIELDKIAAQDFVEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 28) (Compound 28)H-Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAQGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 29) (Compound 29)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQAFVNWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 30) (Compound 30)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAAGPSSGAPPPS-NH₂; (SEQ ID NO: 31) (Compound 31)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 32) (Compound 32)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 33) (Compound 33)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 34) (Compound 34)H-Y-Aib-EGTFISDYSIELD-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-IAQRAFIEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 35) (Compound 35)H-Y-Aib-EGTFISDYS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-ELDKIAQRAFIEWLLAQGPSSGAPPPS-NH_(2;;); (SEQ ID NO: 36) (Compound 36)H-Y-DAla-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 37) (Compound 37)H-Y-DAla-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-NH₂; (SEQ ID NO: 38) (Compound 38)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 39) (Compound 39)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 40) (Compound 40)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS-NH₂;  and or (SEQ ID NO: 41) (Compound 41)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS-NH2.

The invention further provides a pharmaceutical composition comprising aGIP analogue as described herein, or a pharmaceutically acceptable saltor solvate thereof, in admixture with a carrier, preferably apharmaceutically acceptable carrier. The GIP analogue may, for example,be a pharmaceutically acceptable acid addition salt.

The pharmaceutical composition may be formulated as a liquid suitablefor administration by injection or infusion. The pharmaceuticalcomposition may be formulated to cause controlled, e.g., slow release ofsaid GIP analogue.

The invention further provides a therapeutic kit comprising a GIPanalogue as described herein, and a device comprising a GIP analogue asdescribed herein.

The invention further provides a GIP analogue as described herein, or apharmaceutically acceptable salt or solvate thereof, for use in a methodof medical treatment, e.g. for use in the treatment and/or prevention ofa metabolic disorder.

The invention further provides the use of a GIP analogue as describedherein, or a pharmaceutically acceptable salt or solvate thereof, in thepreparation of a medicament for the treatment and/or prevention of ametabolic disorder.

The invention further provides a method of prevention and or/treatmentof a metabolic disorder in a subject, comprising administering a GIPanalogue as described herein, or a pharmaceutically acceptable salt orsolvate thereof, to the subject.

The metabolic disorder may be diabetes or a diabetes related disorder,or obesity or an obesity related disorder. The link between obesity anddiabetes is well known, so these conditions may be but are notnecessarily separate or mutually exclusive.

Diabetes related disorders include insulin resistance, glucoseintolerance, increased fasting glucose, pre-diabetes, type 1 diabetes,type 2 diabetes, gestational diabetes hypertension, dyslipidemia, andcombinations thereof.

Diabetes related disorders also include atherosclerosis,arteriosclerosis, coronary heart disease, peripheral artery disease andstroke; or conditions associated with atherogenic dyslipidemia, bloodfat disorders, elevated blood pressure, hypertension, a prothromboticstate, a proinflammatory state, and bone related disorders such asosteoporosis.

The blood fat disorder may be selected from high triglycerides, low HDLcholesterol, high LDL cholesterol, and plaque buildup in artery walls,or a combination thereof.

The prothrombotic state may be selected from high fibrinogen levels inthe blood and high plasminogen activator inhibitor-1 levels in theblood.

The proinflammatory state may be an elevated C-reactive protein level inthe blood.

Obesity related disorders include obesity linked inflammation, obesitylinked gallbladder disease and obesity induced sleep apnea, or may beassociated with a condition selected from atherogenic dyslipidemia,blood fat disorders, elevated blood pressure, hypertension, aprothrombotic state, and a proinflammatory state, or a combinationthereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Blood glucose levels (A-C) and area under the blood glucosecurves (AUC) (D) in an OGTT in 5-hour fasted mice. The mice wereinjected s.c. with vehicle, the GLP-1 analogue liraglutide (10 nmol/kg),and GIP receptor agonists (compound 12, 13, 17, and 21 at 3-300 nmol/kg)4 hours prior to the oral gavage of glucose (t=0). Data are means±SEM;n=6. Statistical differences vs vehicle: *p<0.05, **p<0.01, ***p<0.001.

FIG. 2: Blood glucose levels (A-D) and area under the blood glucosecurves (AUC) (E) in an OGTT in 5-hour fasted mice. The mice wereinjected s.c. with vehicle and GIP receptor agonists (compounds 12, 18,41, 33, and 35 at 3-300 nmol/kg) 4 hours prior to the oral gavage ofglucose (t=0). Data are means±SEM; n=6. Statistical differences vsvehicle: ***p<0.001.

FIG. 3: Relative body weight changes (delta Δ bodyweight=body weight ateach study day−body weight at day 1) in DIO mice during three weeks oftreatment. Animals were treated once daily with two separate s.c.injections. The first injection was with vehicle 1 or GLP-1 analogueliraglutide (20 nmol/kg). The second injection was with vehicle 2 orCompound 12 (3 and 30 nmol/kg). The GIP agonist was only dosed everythird day of the study (starting on day 1). On other days, GIP agonistwas replaced with vehicle 2. Data are means±SEM; n=8-9. Statisticaldifferences vs vehicle on day 22: ***p<0.001. Statistical difference(p<0.05) between liraglutide and liraglutide co-treated GIP agonist isshown with a line.

FIG. 4: Relative body weight changes (delta Δ body weight=body weight ateach study day−body weight at day 0) in DIO mice during four weeks oftreatment with vehicle, GLP-1 analogue liraglutide, liraglutide+Compound10 or 12 (A), liraglutide+Compound 17 (B), liraglutide+Compound 18 (C),liraglutide+compound 35 (D) or liraglutide+Compound 41 (E). Animals weretreated once daily with two separate s.c. injections. The firstinjection was with vehicle 1 or liraglutide (20 nmol/kg). The secondinjection was with vehicle 2 or GIP agonists (30 and/or 300 nmol/kg).The GIP agonists were only dosed every third day of the study (startingon day 0). On other days, GIP agonists were replaced with vehicle 2.Data are means±SEM; n=9. Statistical differences vs vehicle on day 27:***p<0.001. Statistical differences (p<0.05) between liraglutide andliraglutide co-treated with GIP agonist are shown with lines.

DETAILED DESCRIPTION OF THE INVENTION

Unless otherwise defined herein, scientific and technical terms used inthis application shall have the meanings that are commonly understood bythose of ordinary skill in the art. Generally, nomenclature used inconnection with, and techniques of, chemistry, molecular biology, celland cancer biology, immunology, microbiology, pharmacology, and proteinand nucleic acid chemistry, described herein, are those well known andcommonly used in the art.

Definitions

Unless specified otherwise, the following definitions are provided forspecific terms, which are used in the above written description.

Throughout this specification, the word “comprise” or variations such as“comprises” or “comprising” will be understood to imply the inclusion ofa stated integer (or components) or group of integers (or components),but not the exclusion of any other integer (or components) or group ofintegers (or components).

The singular forms “a,” “an,” and “the” include the plurals unless thecontext clearly dictates otherwise.

The term “including” is used to mean “including but not limited to.”“Including” and “including but not limited to” are used interchangeably.

The terms “patient,” “subject,” and “individual” may be usedinterchangeably and refer to either a human or a non-human animal. Theseterms include mammals such as humans, primates, livestock animals (e.g.,bovines, porcines), companion animals (e.g., canines, felines) androdents (e.g., mice and rats).

The term “solvate” in the context of the present invention refers to acomplex of defined stoichiometry formed between a solute (in casu, apeptide conjugate or pharmaceutically acceptable salt thereof accordingto the invention) and a solvent. The solvent in this connection may, forexample, be water, ethanol or another pharmaceutically acceptable,typically small-molecular organic species, such as, but not limited to,acetic acid or lactic acid. When the solvent in question is water, sucha solvate is normally referred to as a hydrate.

The term “agonist” as employed in the context of the invention refers toa substance (ligand) that activates signaling by the receptor type inquestion. The term “antagonist” as employed in the context of theinvention refers to a substance (ligand) that decreases signaling by thereceptor type in question.

Throughout the description and claims the conventional one-letter andthree-letter codes for natural (or “proteinogenic”) amino acids areused, as well as generally accepted three letter codes for other(non-natural or “non-proteinogenic”) α-amino acids, such as Aib(α-aminoisobutyric acid), Orn (ornithine) and D-Ala (D-alanine). Allamino acid residues in peptides of the invention are preferably of theL-configuration except where explicitly stated.

Among sequences disclosed herein are sequences incorporating an “H—”moiety at the amino terminus (N-terminus) of the sequence, and either an“—OH” moiety or an “—NH₂” moiety at the carboxy terminus (C-terminus) ofthe sequence. In such cases, and unless otherwise indicated, an “H—”moiety at the N-terminus of the sequence in question indicates ahydrogen atom (i.e. R¹═H), corresponding to the presence of a freeprimary or secondary amino group at the N-terminus, while an “—OH” or an“—NH₂” moiety at the C-terminus of the sequence (i.e. R²═OH or NH₂)indicates a carboxy (COOH) group or an amido (CONH₂) group at theC-terminus, respectively.

The compounds of the present invention have GIP biological activity, inparticular in treatment of metabolic diseases such as diabetes andobesity. This can be assessed, e.g., in in vivo assays, in which theblood glucose level or another biological activity is determined after atest animal has been treated or exposed to a GIP analogue. The compoundsof the present invention may be particular effective in improvingglycemic control and reducing body weight when administered togetherwith a GLP-1 receptor agonist to a diabetic patient and/or an overweightor obese subject. The effect obtained with this combination therapy maybe superior to that obtained with the administration of a GLP-1 receptoragonist alone in comparable subjects when given according to acomparable dosing regime. The compounds of the present invention mayalso be capable of improving glycemic control and reducing bodyweightwhen administered alone. The Y1 and Y2 group has a stabilizing effect onthe GIP analogues. Without being bound to any theory it is believed thatgroup comprising the C-terminus part of exendin-4 and GIP compounds hasimpact on the folding of the peptide. In either the treatment of adiabetic subject or an overweight subject, the effect of treating with aGIP analogue of the present invention may be superior to that obtainedwith an equivalent quantity (by mass, or molar ratio) of wild type humanGIP in comparable subjects when given according to a comparable dosingregime, alone or in combination with another anti-diabetic oranti-obesity agent.

Activity in in vitro assays may also be used as a measure of thecompounds' activity. Typically the compounds have activity (i.e. agonistactivity) at the GIP receptor (designated GIP-R). EC₅₀ values may beused as a numerical measure of agonist potency at a given receptor. AnEC₅₀ value is a measure of the concentration of a compound required toachieve half of that compound's maximal activity in a particular assay.In any given assay, the EC₅₀ value of a compound in a given assay may beassessed relative to the EC₅₀ of human GIP. Thus, the ratio of the EC₅₀value of the test compound to the EC₅₀ value of wild type human GIP(EC₅₀[test compound]/EC₅₀[GIP]) at the human GIP receptor may be lessthan 10, less than 5, less than 1, less than 0.1, less than 0.05 or lessthan 0.01. EC₅₀ values may be determined using the human GIP receptorassay described in the Examples below. In such an assay, the compoundsmay, for example, have an EC₅₀ value of 0.001-0.050 nM, 0.001-0.030 nM,0.001-0.020 nM, or 0.001-0.010 nM.

The compounds typically have minimal or no agonist activity at the GLP-1receptor. For example, the ratio of the EC₅₀ value of the test compoundto the EC₅₀ value of the GLP-1 agonist Exendin-4 (EC₅₀[testcompound]/EC₅₀[Ex4]) at the human GIP receptor may be at least about100, at least about 250, at least about 500, at least about 750, atleast about 1000, at least about 5000, or at least about 10,000.(“About” is used here to signify +/−10%.) EC₅₀ values may be determinedusing the human GLP-1 receptor assay described in the Examples below. Insuch an assay, the compounds may, for example, have an EC₅₀ value of atleast 1 nM, at least 3 nM, at least 5 nM or at least 10 nM.

Lipophilic Group

The compound of the invention comprises a residue Ψ, i.e. a residueselected from Lys, Arg, Orn and Cys in which the side chain isconjugated to a lipophilic substituent.

Without wishing to be bound by any particular theory, it is thought thatthe substituent binds plasma proteins (e.g. albumin) in the bloodstream, thus shielding the compounds of the invention from enzymaticdegradation and thereby enhancing the half-life of the compounds. It mayalso modulate the potency of the compound, e.g. with respect to the GIPreceptor.

The substituent is conjugated to the functional group at the distal endof the side chain from the alpha-carbon. The normal ability of the Lys,Arg, Orn or Cys side chain to participate in interactions mediated bythat functional group (e.g. intra- and inter-molecular interactions) maytherefore be reduced or completely eliminated by the presence of thesubstituent. Thus, the overall properties of the compound may berelatively insensitive to changes in the actual amino acid present asresidue Ψ. Consequently, it is believed that any of the residues Lys,Arg, Orn and Cys may be present at any position where L is permitted.However, in certain embodiments, it may be advantageous that the aminoacid component of Ψ is Lys.

Thus, Ψ is a residue of Lys, Arg, Orn or Cys in which the side chain isconjugated to a substituent having the formula —Z¹ or —Z²—Z¹.

—Z¹ is a fatty chain having at a terminus a connection —X— to Ψ or toZ²;

wherein

—X— is a bond, —CO—, —SO—, or —SO₂—;

and, optionally, Z¹ has a polar group at the end of the chain distalfrom connection —X—; said polar group comprising a carboxylic acid or acarboxylic acid bioisostere, a phosphonic acid, or a sulfonic acidgroup;

and wherein —Z²—, if present, is a spacer of formula:

connecting Z¹ to Ψ;wherein:each Y is independently —NH, —NR, —S or —O, where R is alkyl, aprotecting group or forms a linkage to another part of the spacer Z²;each X is independently a bond, CO—, SO—, or SO₂—;with the proviso that when Y is —S, the X to which it is bound is abond;each V is independently a bivalent organic moiety linking Y and X;and n is 1-10.The Group Z¹

Z¹ is a fatty chain having a connection to Ψ or to Z², referred toherein as —X—. —X— may be, for example, a bond, acyl (—CO—), sulfinyl(—SO—), or sulfonyl (—SO₂—). When Z¹ is bound directly to Ψ, that is,when Z² is not present, preferably —X— is acyl (—CO—), sulfinyl (—SO—),or sulfonyl (—SO₂—). Most preferably, —X— is acyl (—CO—).

Z¹ may further have a polar group, said polar group being located at theend of the chain distal from the connection —X—. In other words, theconnection is located at the w-position with respect to the polar group.The polar group may be bound directly to the terminus of the fattychain, or may be bound via a linker.

Preferably, the polar group is an acidic or weakly acid group, forexample a carboxylic acid or a carboxylic acid bioisostere, aphosphonate, or a sulfonate. The polar group may have a pK_(a) ofbetween −2 and 12 in water, more preferably between 1 and 7, morepreferably between 3 and 6. Certain preferred polar groups have a pK_(a)of between 4 and 5.

For example, and not by way of limitation, the polar group may comprisea carboxylic acid (—COOH) or a carboxylic acid bioisostere, a phosphonicacid (—P(O)(OH)₂), or a sulfonic acid (—SO₂OH) group.

Preferably the polar group, if present, comprises a carboxylic acid orcarboxylic acid bioisostere. Suitable carboxylic acid bioisosteres areknown in the art. Preferably the bioisostere has a proton having apK_(a) similar to the corresponding carboxylic acid. Examples ofsuitable bioisoteres may include, not by way of limitation, tetrazole,acylsulfomides, acylhydroxylamine, and squaric acid derivatives, asshown below (--- indicates the point of attachment):

Fatty chain as used herein refers to a moiety comprising a chain ofcarbon atoms, the carbon atoms being predominantly substituted withhydrogen or hydrogen-like atoms, for example, a hydrocarbon chain. Suchfatty chains are often referred to as lipophilic, although it will beappreciated that substitution may alter the lipophilic properties of theoverall molecule.

The fatty chain may by aliphatic. It may be entirely saturated or mayinclude one or more double or triple bonds. Each double bond, ifpresent, may be in the E or Zconfiguration. The fatty chain may alsohave one or more cycloalkylene or heterocycloalkylene moieties in itslength, and additionally or alternatively may have one or more aryleneor heteroarylene moieties in its length. For example, the fatty chainmay incorporate a phenylene or piperazinylene moiety in its length as,for example, shown below (wherein --- represents the points ofattachment within the chain).

The fatty chain may be derived from a fatty acid, for example, it may bederived from a medium-chain fatty acid (MCFA) with an aliphatic tail of6-12 carbon atoms, a long-chain fatty acid (LCFA) with an aliphatic tailof 13-21 carbon atoms, or a very long-chain fatty acid (LCFA) with analiphatic tail of 22 carbon atoms or more. Examples of linear saturatedfatty acids from which suitable fatty chains may be derived includetridecylic (tridecanoic) acid, myristic (tetradecanoic) acid,pentadecylic (pentadecanoic) acid, palmitic (hexadecanoic) acid, andmargaric (heptadecanoic) acid. Examples of linear unsaturated fattyacids from which suitable fatty chains may be derived includemyristoleic acid, palmitoleic acid, sapienic acid and oleic acid.

The fatty chain may be connected to Ψ or to Z² by an amide linkage, asulfinamide linkage, a sulfonamide linkage, or by an ester linkage, orby an ether, thioether or amine linkage. Accordingly, the fatty chainmay have, a bond to Ψ or to Z² or an acyl (—CO—), sulfinyl (—SO—), orsulfonyl (—SO₂—) group. Preferably, the fatty chain has a terminushaving an acyl (—CO—) group and is connected to Ψ or Z² by an amide orester linkage.

In some embodiments, Z¹ is a group of formula:A-B-Alk-X—whereinA is hydrogen or a carboxylic acid, a carboxylic acid bioisostere, aphosphonic acid, or a sulfonic acid group;B is a bond or a linker;X is a bond, acyl (—CO—), sulfinyl (—SO—), or sulfonyl (—SO₂—); andAlk is a fatty chain that may be optionally substituted with one or moresubstituents. The fatty chain is preferably 6 to 28 carbon atoms inlength (e.g. a C₆₋₂₈alkylene), more preferably, 12 to 26 carbons inlength (e.g. a C₁₂₋₂₆alkylene), more preferably, 16 to 22 carbons inlength (e.g. C₁₆₋₂₂alkylene), and may be saturated or unsaturated.Preferably, Alk is saturated, that is, preferably Alk is alkylene.

Optional substituents on the fatty chain may be independently selectedfrom fluoro, C₁₋₄alkyl, preferably methyl; trifluoromethyl,hydroxymethyl, amino, hydroxyl, C₁₋₄alkoxy, preferably methoxy; oxo, andcarboxyl, and may be independently located at any point along the chain.In some embodiments, each optional substituent is selected from fluoro,methyl, and hydroxyl. Where more than one substituent is present,substituents may be the same or different. Preferably, the number ofsubstituents is 0 to 3; more preferably the fatty chain isunsubstituted.

B may be a bond or a linker. When B is a linker, it may be acycloalkylene, heterocycloalkylene, C₆arylene, or C₅₋₆heteroarylene, orC₆arylene-O— or C₅₋₆heteroarylene-O—.

When B is phenylene it may, for example, be selected from 1,2-phenylene,1,3-phenylene, 1,4-phenylene, preferably 1,4-phenylene (so that A-B- isa 4-benzoic acid substituent or 4-benzoic acid bioisostere). When B isphenylene-O—, it may, for example, be selected from 1,2-phenylene-O—,1,3-phenylene-O—, 1,4-phenylene-O—, preferably 1,4-phenylene-O. Eachphenylene of B may be optionally substituted with one or moresubstituents selected from fluoro, methyl, trifluoromethyl, amino,hydroxyl, and C₁₋₄alkoxy, preferably methoxy. It will be appreciatedthat substituent identity and position may be selected to subtly alterthe pK_(a) of the polar group. Suitable inductively or mesomericallyelectron-withdrawing or donating groups and their positional effects areknown in the art. In some embodiments, B may be C₅₋₆heteroarylene, forexample, pyridinylene or thiofuranylene, and may be optionallysubstituted as described.

For example, in some embodiments, A-B— may be selected from:

Preferably, A is H— or HOOC— and B is a bond.

It will be understood that when A is hydrogen, B is a bond and Alk isunsubstituted alkylene, A-B-Alk- is an alkyl chain of formulaH₃C—(CH₂)_(n)—.

In some embodiments, Z¹ is an acyl group of formula:A-B-Alk-(CO)—or a sulfonyl group of formula:A-B-Alk-(SO₂)—.

Preferably, Z¹ is an acyl group of formula:A-B-alkylene-(CO)—where A and B are as defined above.

In some embodiments, A is —COOH and B is a bond. Accordingly, certainpreferred Z¹ are derived from long-chain saturated α,ω-dicarboxylicacids of formula HOOC—(CH₂)₁₂₋₂₂—COOH, preferably, long-chain saturatedα,ω-dicarboxylic acids having an even number of carbon atoms in thealiphatic chain. In some other embodiments, A is H and B is a bond.Accordingly, certain preferred Z¹ are derived from long-chain saturatedcarboxylic acids of formula HOOC—(CH₂)₁₂₋₂₂—CH₃, preferably, long-chainsaturated carboxylic acids having an even number of carbon atoms in thealiphatic chain.

For example, and not by way of limitation, Z¹ may be:

A-B—C₁₆₋₂₀alkylene-(CO)— wherein A is H or —COOH and B is a bond, forexample:

17-carboxy-heptadecanoyl HOOC—(CH₂)₁₆—(CO)—;

19-carboxy-nonadecanoyl HOOC—(CH₂)₁₈—(CO)—;

Octadecanoyl H₃C—(CH₂)₁₆—(CO)—;

Eicosanoyl H₃C—(CH₂)₁₈—(CO)—;

The carboxylic acid group, if present, may be replaced by a bioisotereas detailed herein.

The group Z²

Z² is an optional spacer that connects Z¹ to the side chain of the aminoacid component of Ψ. At its most general, Z², if present, is a spacerbound at one terminus by Y, which may be a nitrogen, oxygen or sulfuratom, and at the other terminus by X, which may be a bond or an acyl(—CO—), sulfinyl (—SO—), sulfonyl (—SO₂—) or absent. Accordingly, Z² maybe a spacer of formula (--- indicate points of attachment):

wherein:Y may be —NH, —NR, —S or —O, where R may be alkyl, a protecting group ormay form a linkage to another part of the spacer, with the remainingvalency forming a linkage to Z¹;X may be a bond, CO—, SO—, or SO₂—, with the remaining valency forming alinkage to the side chain of the amino acid component of Ψ;V is a bivalent organic moiety linking Y and X;and n may be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. Where n is 2 or more, eachY, V, and X is independent of every other Y, V, and X.

Accordingly, Z² may be bound at each side by amide, sulfinamide,sulfonamide, or ester linkages or by amino, ether, or thioether linkagesdepending upon the nature of Y and X and the corresponding linkinggroups on Z¹ and the side chain. Where n is 2 or greater, each V mayalso be bound to each adjacent V by linkages as described. Preferably,linkages are amides, esters or sulfonamides, most preferably amides.Accordingly, in some embodiments, each Y is —NH or —NR and each X is CO—or SO₂—. Most preferably, —X— is acyl (—CO—).

In some embodiments, Z² is a spacer of formula —S_(A)—, —S_(B)—,—S_(A)—S_(B)— or —S_(B)—S_(A)—, wherein S_(A) and S_(B) are as definedbelow.

In some embodiments, Z² is selected from —S_(A)— or —S_(B)—S_(A)—, thatis, [side chain]-Z²Z¹ is [side chain]-S_(A)—Z¹ or [sidechain]-S_(B)-S_(A)—Z¹.

The Group S_(A)

S_(A) may be a single amino acid residue or a residue of an amino acidderivative, especially an amino acid derivative residue having asulfinyl or sulfonyl in place of the carboxy moiety at the C terminus.Additionally or alternatively, the single amino acid residue may have anoxygen or sulfur atom in place of the nitrogen atom at the N terminus.

S_(A) may be or may comprise a nitrogen-containing heterocycle, saidnitrogen-containing heterocycle being bound within the lipophilic groupat one end via a bond, a carboxy, a sulfinyl, or a sulfonyl group and atthe other via a ring nitrogen atom. For example, S_(A) may comprise apiperazine ring.

Suitably, S_(A) is a 5-8-membered heterocycle having 1 or 2 nitrogenatoms and substituted with an X group, where X is a bond, CO—, SO—, orSO₂—, and where L, if present, is C₁₋₄alkylene (— denotes a point ofattachment within the lipophilic group).

Preferably, S_(A) is a 6-membered heterocycle having 1 or 2 nitrogenatoms, preferably 2, and substituted with a —CH₂CO—, —CH₂SO—, or—CH₂SO₂— group.

For example, S_(A) may be:

For example, S_(A) may be:

(referred to herein as piperazine-1-yl-acetyl).

Preferably, S_(A) is a single amino acid residue orpiperazine-1-yl-acetyl. More preferably S_(A) is a single amino acidresidue.

In some embodiments, the amino acid may be selected from γ-Glu, α-Glu,α-Asp, β-Asp, Ala, β-Ala (3-aminopropanoic acid), Dapa(2,3-diaminopropanoic acid), Dab (2,4-diaminobutanoic acid), and Gaba(4-aminobutanoic acid). It will be understood that where more than onecarboxylic acid or amino moiety is present, connection may be at anymoiety as appropriate. Any carboxylic acid or amino resides not boundwithin the residue may be free, that is, present as a free carboxylicacid or primary amine, or may be derivatised. Suitable derivatisation isknown in the art. For example, carboxylic acid moieties may be presentin S_(A) amino acid residues as esters, for example, as methyl esters.Amino moieties may be present as alkylated amines, for example,methylated, or may be protected as amide or carbamate moieties. Othersuitable amino acids include β-Ala (3-aminopropanoic acid) and Gaba(4-aminobutanoic acid) and similar w amino acids.

It will be understood that amino acids may be D or L, or a racemic orenantioenriched mixture. In some embodiments, the amino acid is anL-amino acid. In some embodiments, the amino acid is a D-amino acid.

In some preferred embodiments, S_(A) has a carboxylic acid substituent,with γ-Glu, α-Glu, α-Asp, and β-Asp, and sulfinyl and sulfonylderivatives thereof, being preferred. Accordingly, in some embodiments,the amino acid residue is:

where —X— is —CO—, —SO—, —SO₂—, preferably —CO—, and a is 1 or 2,preferably 2. In some embodiments, the carboxylic acid is an ester, andthe amino acid residue is:

where —X— is —CO—, —SO—, —SO₂—, preferably —CO—, and a is 1 or 2,preferably 2, and R is C₁₋₄alkyl or C₆aryl. Preferably R is C₁₋₄alkyl,preferably methyl or ethyl, more preferably ethyl.

A preferred S_(A) group bearing a carboxylic acid is γ-Glu.

Preferably, S_(A) is selected from Dapa or γ-Glu. Most preferably, S_(A)is γ-Glu.

The Group S_(B)

S_(B) may be a linker of general formula:

wherein P_(U) is a polymeric unit and n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or10. One terminus of the linker S_(B) is an —NH, —NR, —S or —O, wherein Rmay be alkyl, a protecting group or may form a linkage to another partof the polymeric unit; while the other is a bond or CO—, SO— or SO₂—.Accordingly, each polymeric unit P_(U) may be bound at each side byamide, sulfinamide, sulfonamide, or ester linkages or by amino, ether,or thioether linkages depending upon the nature of Y and X and thecorresponding linking groups on Z¹, S_(A), and Lys.

In some embodiments, each P_(U) may be independently a unit of formula:

wherein:Y may be —NH, —NR, —S or —O, wherein R may be alkyl, a protecting groupor may form a linkage to another part of the spacer, with the remainingvalency forming a linkage to Z¹;X may be a bond, CO—, SO—, or SO₂—, with the remaining valency forming alinkage to the Ψ side chain;and V is a bivalent organic moiety linking Y and X.

In some embodiments, V is the α-carbon of a natural or unnatural aminoacid, that is V is —CHR^(AA)—, wherein R^(AA) is an amino acid sidechain; or V is an optionally substituted C₁₋₆alkylene, or V is a chaincomprising one or more units of ethylene glycol in series, also known asPEG chain, for example, —CH₂CH₂—(OCH₂CH₂)_(m)—O—(CH₂)_(p)—, where m is0, 1, 2, 3, 4, or 5, and p is 1, 2, 3, 4, or 5; when X is CO—, p ispreferably 1, 3, 4, or 5. Optional alkylene substituents include fluoro,methyl, hydroxy, hydroxymethyl, and amino.

Preferred P_(U) units include:

(i). Single amino acid residues: P_(U) ^(i);

(ii). Dipeptide residues: P_(U) ^(ii); and

(iii). Amino-(PEG)_(m)-carboxylic acid residues: P_(U) ^(iii),

and may be present in any combination or order. For example, S_(B) maycomprise one or more of each of P_(U) ^(i), P_(U) ^(ii), and P_(U)^(iii) in any order, or may comprise one or more units of P_(U) ^(i),P_(U) ^(ii), and P_(U) ^(iii) only, or one of more units selected fromP_(U) ^(i) and P_(U) ^(ii), P_(U) ^(i) and P_(U) ^(iii), or P_(U) ^(ii)and P_(U) ^(iii).(i). P_(U) ^(i) Single Amino Acid Residues

Each P_(U) ^(i) may be independently selected from any natural orunnatural amino acid residue and, for example, may be selected from Gly,Pro, Ala, Val, Leu, Ile, Met, Cys, Phe, Tyr, Trp, His, Lys, Arg, Gln,Asn, α-Glu, γ-Glu, Asp, Ser Thr, Dapa, Gaba, Aib, β-Ala,5-aminopentanoyl, 6-aminohexanoyl, 7-aminoheptanoyl, 8-aminooctanoyl,9-aminononanoyl, and 10-aminodecanoyl. Preferably, P_(U) ^(i) amino acidresidues are selected from Gly, Ser, Ala, Thr, and Cys, more preferablyfrom Gly and Ser.

In some embodiments, S_(B) is —(P_(U) ^(i))_(n)—, wherein n is 1 to 8,more preferably 5 to 7, most preferably 6. In some preferredembodiments, S_(B) is —(P_(U) ^(i))_(n)—, n is 6 and each P_(U) ^(i) isindependently selected from Gly or Ser, with a preferred sequence being-Gly-Ser-Gly-Ser-Gly-Gly-.

(ii). P_(U) ^(ii) Dipeptide Residues

Each P_(U) ^(ii) may be independently selected from any dipeptideresidue comprising two natural or unnatural amino acid residues bound byan amide linkage. Preferred P_(U) ^(ii) dipeptide residues includeGly-Gly, Gly-Ser, Ser-Gly, Gly-Ala, Ala-Gly, and Ala-Ala, morepreferably Gly-Ser and Gly-Gly.

In some embodiments, S_(B) is —(P_(U) ^(ii))_(n)—, wherein n is 2 to 4,more preferably 3, and each P_(U) ^(ii) is independently selected fromGly-Ser and Gly-Gly. In some preferred embodiments S_(B) is —(P_(U)^(ii))_(n)—, n is 3 and each P_(U) ^(ii) is independently selected fromGly-Ser and Gly-Gly, with a preferred sequence being-(Gly-Ser)-(Gly-Ser)-(Gly-Gly) (SEQ ID NO: 86).

Amino acids having stereogenic centres within P_(U) ^(i) and P_(U) ^(ii)may be racemic, enantioenriched, or enantiopure. In some embodiments,the or each amino acid is independently an L-amino acid. In someembodiments, the or each amino acid is independently a D-amino acid.

(iii). P_(U) ^(iii) amino-(PEG)_(m)-carboxylic Acid Residues

Each P_(U) ^(iii) may be independently a residue of general formula:

wherein m is 0, 1, 2, 3, 4, or 5, preferably 1 or 2, and p is 1, 3, 4,or 5, preferably 1. In some embodiments, m is 1 and p is 1, that is,P_(U) ^(iii) is a residue of 8-amino-3,6-dioxaoctanoic acid (also knownas {2-[2-aminoethoxy]ethoxy}acetic acid and H₂N-PEG₃-COOH). This residueis referred to herein as -PEG₃-.

Other, longer, PEG chains are also known in the art. For example,11-amino-3,6,9-trioxaundecanoic acid (also known as H₂N-PEG₄-COOH or-PEG₄-).

In some embodiments, S_(B) is —(P_(U) ^(iii))_(n)—, wherein n is 1 to 3,more preferably 2.

Most preferably, S_(B) is -PEG₃-PEG₃-.

Preferred Combinations

It will be understood that the above preferences may be independentlycombined to give preferred —Z¹ and —Z²—Z¹ moieties.

Some preferred —Z¹ and —Z²—Z¹ moieties are shown below (in each case,--- indicates the point of attachment to the side chain of the aminoacid component of Ψ:

The skilled person will be well aware of suitable techniques forpreparing the compounds employed in the context of the invention. Forexamples of suitable chemistry, see, e.g., WO98/08871, WO00/55184,WO00/55119, Madsen et al. (J. Med. Chem. 2007, 50, 6126-32), and Knudsenet al. 2000 (J. Med Chem. 43, 1664-1669).

Clinical Utility

The GIP analogue compounds employed in the context of the invention mayprovide an attractive treatment option for metabolic diseases includingobesity, diabetes mellitus (diabetes), obesity-related disorders, anddiabetes-related disorders. The GIP analogue compounds of the presentinvention may be particular effective in improving glycaemic control andreducing body weight when they are administered in combination with aGLP-1 receptor agonist (as part of the same pharmaceutical formulationor as separate formulations). Glucagon-like peptide-1 receptor agonistsalso known as GLP-1 receptor agonists or incretin mimetics are agonistsof the GLP-1 receptor. One of their advantages over older insulinsecretagogues, such as sulfonylureas or meglitinides, is that they havea lower risk of causing hypoglycemia.

Examples of GLP-1 agonists include but are not limited to exenatide(Byetta®/Bydureon®), liraglutide (Victoza®), semaglutide, lixisenatide(Lyxumia®), albiglutide (Tanzeum®) and Taspoglutide.

Diabetes comprises a group of metabolic diseases characterized byhyperglycemia resulting from defects in insulin secretion, insulinaction, or both. Diabetes is classified into type 1 diabetes, type 2diabetes and gestational diabetes on the basis on pathogeniccharacteristics. Type 1 diabetes accounts for 5-10% of all diabetescases and is caused by auto-immune destruction of insulin-secretingpancreatic β-cells. Acute signs of diabetes include excessive urineproduction, resulting compensatory thirst and increased fluid intake,blurred vision, unexplained weight loss, lethargy, and changes in energymetabolism. However, in type 2 diabetes symptoms are often not severe ormay be absent. The chronic hyperglycemia of diabetes is associated withlong-term damage, dysfunction, and failure of various organs, notablythe eyes, kidneys, nerves, heart and blood vessels.

Type 2 diabetes accounts for 90-95% of diabetes cases and is a result ofa complex set of metabolic disorders. However, symptoms are often notsevere or may be absent. Type 2 diabetes is the consequence ofendogenous insulin production becoming insufficient to maintain plasmaglucose levels below diagnostic thresholds.

Gestational diabetes refers to any degree of glucose intoleranceidentified during pregnancy.

Pre-diabetes includes impaired fasting glucose and impaired glucosetolerance and refers to those states that occur when blood glucoselevels are elevated but below the levels that are established for theclinical diagnosis for diabetes.

A large proportion of people with type 2 diabetes and pre-diabetes areat increased risk of morbidity and mortality due to the high prevalenceof additional metabolic risk factors, including abdominal obesity(excessive fat tissue around the abdominal internal organs), atherogenicdyslipidemia (blood fat disorders including high triglycerides, low HDLcholesterol and/or high LDL cholesterol, which foster plaque buildup inartery walls), elevated blood pressure (hypertension) a prothromboticstate (e.g. high fibrinogen or plasminogen activator inhibitor- 1 in theblood), and/or a proinflammatory state (e.g., elevated C-reactiveprotein in the blood).

Conversely, obesity confers an increased risk of developingpre-diabetes, type 2 diabetes as well as, e.g., certain types of cancer,obstructive sleep apnea and gall-bladder disease. Dyslipidemia isassociated with increased risk of cardiovascular disease. High DensityLipoprotein (HDL) is of clinical importance since an inverse correlationexists between plasma HDL concentrations and risk of atheroscleroticdisease. The majority of cholesterol stored in atherosclerotic plaquesoriginates from LDL and hence an elevated concentration of Low DensityLipoproteins (LDL) is closely associated with atherosclerosis. TheHDL/LDL ratio is a clinical risk indictor for atherosclerosis andcoronary atherosclerosis in particular.

The GIP analogues of the present invention may be used as pharmaceuticalagents for preventing weight gain, promoting weight loss, reducingexcess body weight or treating obesity (e.g., by control of appetite,feeding, food intake, calorie intake, and/or energy expenditure andlipolysis), including morbid obesity, as well as associated diseases andhealth conditions including but not limited to obesity linkedinflammation, obesity linked gallbladder disease and obesity inducedsleep apnea. The GIP analogues employed in the context of the inventionmay also be used for treatment of insulin resistance, glucoseintolerance, pre-diabetes, increased fasting glucose, type 2 diabetes,hypertension, dyslipidemia (or a combination of these metabolic riskfactors), atherosclerosis, arteriosclerosis, coronary heart disease,peripheral artery disease and stroke. These are all conditions which maybe associated with obesity. However, the effects of the compoundsemployed in the context of the invention on these conditions may bemediated in whole or in part via an effect on body weight, or may beindependent thereof.

The GIP analogues of the present invention may thus be used for thetreatment and/or prevention of any of the diseases, disorders, orconditions described herein, including insulin resistance, glucoseintolerance, increased fasting glucose, pre-diabetes, type 1 diabetes,type 2 diabetes, gestational diabetes hypertension, dyslipidemia, or acombination thereof. In certain embodiments, the diabetes relateddisorder is selected from atherosclerosis, arteriosclerosis, coronaryheart disease, peripheral artery disease and stroke; or associated witha condition selected from atherogenic dyslipidemia, blood fat disorders,elevated blood pressure, hypertension, a prothrombotic state, andproinflammatory state, or a combination thereof. In certain embodiments,the blood fat disorder is selected from high triglycerides, low HDLcholesterol, high LDL cholesterol, plaque buildup in artery walls, or acombination thereof. In certain embodiments, the prothrombotic state isselected from high fibrinogen levels in the blood and high plasminogenactivator inhibitor-1 levels in the blood. In certain embodiments, theproinflammatory state is an elevated C-reactive protein level in theblood. In certain embodiments, the obesity related disorder is selectedfrom obesity linked inflammation, obesity linked gallbladder disease andobesity induced sleep apnea.

The GIP analogues of the present invention may also be used for thetreatment and/or prevention of any of the diseases, disorders, orconditions associated with diabetes related osteoporosis includingincreased risk of bone fractures (Khazai N. B. et al, 2009, CurrentOpinion in Endocrinology, Diabetes and Obesity, vol. 16, no. 6,435-445). The increase in fracture risk is likely to be related toimpaired bone quality rather than to bone mineral density. The relatedmechanisms, due at least in part to hyperglycemia, neuropathy, andhigher incidence of hypovitaminosis D, are not yet fully understood(Takiishi T et al, 2010, Endocrinology and Metabolism Clinics of NorthAmerica, vol. 39, no. 2, 419-446).

In some embodiments, the invention also provides a therapeutic kitcomprising a GIP analogue (e.g., GIP agonist compound) of the presentinvention, optionally in combination with a pharmaceutically acceptablecarrier. In some embodiments, the invention provides a device comprisinga GIP analogue of the invention for delivery of the GIP analogue to asubject.

Pharmaceutical Compositions

The GIP analogues (e.g., GIP agonist compounds) of the presentinvention, or salts or solvates thereof, may be formulated aspharmaceutical compositions prepared for storage or administration,which typically comprise a therapeutically effective amount of acompound employed in the context of the invention, or a salt or solvatethereof, in a pharmaceutically acceptable carrier. In some embodiments,the pharmaceutical composition is formulated as a liquid suitable foradministration by injection or infusion, or which is formulated to causeslow release of the GIP analogue.

The therapeutically effective amount of a compound of the presentinvention will depend, e.g., on the route of administration, the type ofmammal being treated, and the physical characteristics of the specificmammal under consideration. These factors and their relationship todetermining this amount are well known to skilled practitioners in themedical arts. This amount and the method of administration can betailored to achieve optimal efficacy, and may depend on such factors asweight, diet, concurrent medication and other factors, well known tothose skilled in the medical arts. The dosage sizes and dosing regimenmost appropriate for human use may be guided by the results obtained bythe present invention, and may be confirmed in properly designedclinical trials.

An effective dosage and treatment protocol may be determined byconventional means, starting with a low dose in laboratory animals andthen increasing the dosage while monitoring the effects, andsystematically varying the dosage regimen as well. Numerous factors maybe taken into consideration by a clinician when determining an optimaldosage for a given subject. Such considerations are known to the skilledperson. The term “pharmaceutically acceptable carrier” includes any ofthe standard pharmaceutical carriers. Pharmaceutically acceptablecarriers for therapeutic use are well known in the pharmaceutical art,and are described, for example, in Remington's Pharmaceutical Sciences,Mack Publishing Co. (A. R. Gennaro edit. 1985). For example, sterilesaline and phosphate-buffered saline at slightly acidic or physiologicalpH may be used. Suitable pH buffering agents may be, e.g., phosphate,citrate, acetate, lactate, maleate, tris/hydroxymethyl)aminomethane(TRIS), N-Tris(hydroxymethyl)methyl-3-aminopropanesulphonic acid (TAPS),ammonium bicarbonate, diethanolamine, histidine, which in certainembodiments is a preferred buffer, arginine, lysine, or acetate ormixtures thereof. The term further encompasses any agents listed in theUS Pharmacopeia for use in animals, including humans.

The term “pharmaceutically acceptable salt” refers to a salt of thecompound. Salts include pharmaceutically acceptable salts, such as,e.g., acid addition salts and basic salts. Examples of acid additionsalts include hydrochloride salts, citrate salts and acetate salts.Examples of basic salts include salts where the cation is selected fromalkali metals, such as sodium and potassium, alkaline earth metals suchas calcium, and ammonium ions ⁺N(R³)₃(R⁴), where R³ and R⁴ independentlydesignate optionally substituted C₁₋₆-alkyl, optionally substitutedC₂₋₆-alkenyl, optionally substituted aryl, or optionally substitutedheteroaryl. Other examples of pharmaceutically acceptable salts aredescribed in “Remington's Pharmaceutical Sciences”, 17th edition. Ed.Alfonso R. Gennaro (Ed.), Mark Publishing Company, Easton, Pa., U.S.A.,1985 and more recent editions, and in the Encyclopaedia ofPharmaceutical Technology.

“Treatment” is an approach for obtaining beneficial or desired clinicalresults. For purposes of this invention, beneficial or desired clinicalresults include, but are not limited to, alleviation of symptoms,diminishment of extent of disease, stabilized (i.e., not worsening)state of disease, delay or slowing of disease progression, ameliorationor palliation of the disease state, and remission (whether partial ortotal), whether detectable or undetectable. “Treatment” may also meanprolonging survival as compared to expected survival if not receivingtreatment. “Treatment” is an intervention performed with the intentionof preventing the development or altering the pathology of a disorder.Accordingly, “treatment” refers to both therapeutic treatment andprophylactic or preventative measures in certain embodiments. Those inneed of treatment include those already with the disorder as well asthose in which the disorder is to be prevented. By treatment is meantinhibiting or reducing an increase in pathology or symptoms (e.g. weightgain, hyperglycemia) when compared to the absence of treatment, and isnot necessarily meant to imply complete cessation of the relevantcondition.

The pharmaceutical compositions of the invention may be in unit dosageform. In such form, the composition is divided into unit dosescontaining appropriate quantities of the active component. The unitdosage form can be a packaged preparation, the package containingdiscrete quantities of the preparations, for example, packeted tablets,capsules, and powders in vials or ampoules. The unit dosage form canalso be a capsule, cachet, or tablet itself, or it can be theappropriate number of any of these packaged forms. It may be provided insingle dose injectable form, for example in the form of an injectionpen. Compositions may be formulated for any suitable route and means ofadministration. Pharmaceutically acceptable carriers or diluents includethose used in formulations suitable for oral, rectal, nasal orparenteral (including subcutaneous, intramuscular, intravenous,intradermal, and transdermal) administration. The formulations mayconveniently be presented in unit dosage form and may be prepared by anyof the methods well known in the art of pharmacy. Subcutaneous ortransdermal modes of administration may be particularly suitable forcertain of the compounds described herein.

Combination Therapy

In certain embodiments, a GIP-analogue employed in the context of theinvention may be administered as part of a combination therapy with atleast one other agent for treatment of diabetes, obesity, dyslipidemia,or hypertension.

In such cases, the at least two active agents may be given together orseparately, and as part of the same pharmaceutical formulation or asseparate formulations. Thus, the GIP analogue employed in the context ofthe invention (or the salt or solvate thereof) may be used incombination with an antidiabetic agent including but not limited to aglucagon-like peptide receptor 1 agonist, metformin, a sulfonylurea, aglinide, a DPP-IV inhibitor, a glitazone, or insulin. In certainembodiments, the compound or salt or solvate thereof is used incombination with insulin, DPP-IV inhibitor, sulfonylurea or metformin,particularly sulfonylurea or metformin, for achieving adequate glycemiccontrol. In certain preferred embodiments, the compound or salt orsolvate thereof is used in combination with insulin or an insulinanalogue for achieving adequate glycemic control. Examples of insulinanalogues include but are not limited to LANTUS®, NOVORAPID®, HUMALOG®,NOVOMIX®, ACTRAPHANE HM®, LEVEMIR®, and APIDRA®.

In certain embodiments, the GIP analogue or salt or solvate thereof mayfurther be used in combination with one or more of an anti-obesityagent, including but not limited to a glucagon-like peptide receptor 1agonist, peptide YY or analogue thereof, cannabinoid receptor 1antagonist, lipase inhibitor, melanocortin receptor 4 agonist, ormelanin concentrating hormone receptor 1 antagonist.

In certain embodiments, the GIP analogue or salt or solvate thereof maybe used in combination with an anti-hypertension agent, including butnot limited to an angiotensin-converting enzyme inhibitor, angiotensinII receptor blocker, diuretics, beta-blocker, or calcium channelblocker.

In certain embodiments, the GIP analogue or salt thereof may be used incombination with an anti-dyslipidemia agent, including but not limitedto a statin, a fibrate, a niacin and/or a cholesterol absorptioninhibitor.

Synthesis of Compounds of the Invention

A nucleic acid molecule may encode the amino acid sequence of any ofFormula I to III or a precursor thereof. The amino acid sequence encodedcan be regarded as a precursor of a compound of the invention.

Typically, such nucleic acid sequences will be provided as expressionconstructs wherein the encoding nucleic acid is in functional linkagewith appropriate control sequences to direct its expression. Theexpression construct may be provided in the context of a host cellcapable of expressing (and optionally also secreting) the amio acidprecursor, or ina cell-free expression system.

The invention provides a method of producing a GIP analogue of theinvention, the method comprising expressing an amino acid precursor ofthe GIP analogue and modifying the precursor to provide the GIPanalogue. The modification may comprise chemical modification of a Lys,Arg or Cys residue present at position 17 to introduce the lipophilicmoiety, modification of the N- or C-terminus, and/or modification of anyother amino acid side chains in the molecule (e.g. to introduce anon-naturally occurring amino acid residue).

The compounds of the invention may also be manufactured by standardpeptide synthetic methods, e.g. by standard solid-phase or liquid-phasemethodology, either stepwise or by fragment assembly, and isolating andpurifying the final peptide compound product, or by any combinations ofrecombinant and synthetic methods.

It may be preferable to synthesize the peptide compounds of theinvention by means of solid-phase or liquid-phase peptide synthesis. Inthis context, reference may be made to WO 98/11125 or, inter alia,Fields, G. B. et al., “Principles and Practice of Solid-Phase PeptideSynthesis”; in: Synthetic Peptides, Gregory A. Grant (ed.), OxfordUniversity Press (2nd edition, 2002) and the synthesis examples herein.

EXAMPLES

The following examples demonstrate certain embodiments of the presentinvention. However, it is to be understood that these examples neitherpurport nor are they intended to be wholly definitive as to conditionsand scope of this invention. The examples were carried out usingstandard techniques, which are well known and routine to those of skillin the art, except where otherwise described in detail. The followingexamples are presented for illustrative purposes only, and should not beconstrued in any way as limiting the scope of this invention.

Disclosed are GIP analogues that exhibit signaling selectivity, andmethods for screening these compounds. Signaling selectivity may be, forexample, preferential pathway activation or preferential pathwayinhibition, or both. The analogue, administered alone or in combinationwith a GLP-1 agonist, may be useful for the treatment and/or preventionof diseases or conditions caused or characterized by excess body weight,including, but not limited to, obesity, morbid obesity, obesity linkedinflammation, obesity linked gallbladder disease, obesity induced sleepapnea, metabolic syndrome, pre-diabetes, insulin resistance, glucoseintolerance, type 2 diabetes, type I diabetes, hypertension, atherogenicdyslipidaemia, atherosclerosis, arteriosclerosis, coronary heartdisease, peripheral artery disease, and stroke or microvascular disease.

While some embodiments of the invention have been described by way ofillustration, it will be apparent that the invention can be put intopractice with many different modifications, variations and adaptations,and with the use of numerous equivalents or alternative solutions thatare within the scope of persons skilled in the art, without departingfrom the spirit of the invention or exceeding the scope of the claims.

All publications, patents, and patent applications referred to hereinare herein incorporated by reference in their entirety to the sameextent as if each individual publication, patent or patent applicationwas specifically and individually indicated to be incorporated byreference in its entirety.

The methods used in the instant invention are described below, exceptwhere expressly indicated otherwise.

Example 1

General Synthesis of Acylated GIP Analogues

Solid phase peptide synthesis was performed on a CEM Liberty PeptideSynthesizer using standard Fmoc chemistry. TentaGel S Ram resin (1 g;0.25 mmol/g) was swelled in NMP (10 ml) prior to use and transferredbetween tube and reaction vessel using DCM and NMP.

Coupling

An Fmoc-amino acid in DMF/DCM (2:1; 0.2 M; 5 ml) was added to the resinin a CEM Discover microwave unit together with HATU/DMF or COMU/DMF (0.5M; 2 ml) and DIPEA-DMF/DCM (2:1) (2.0 M; 1 ml). The coupling mixture washeated to 75° C. for 5 min while nitrogen was bubbled through themixture. The resin was then washed with DMF (4×10 ml).

Deprotection

Piperidine/DMF (20%; 10 ml) was added to the resin for initialdeprotection and the mixture was heated by microwaves (30 sec; 40° C.).The reaction vessel was drained and a second portion of piperidine/NMP(20%; 10 ml) was added and heated (75° C.; 3 min.) again. The resin wasthen washed with DMF (6×10 ml).

Side Chain Acylation

Fmoc-Lys(ivDde)-OH or alternatively another amino acid with anorthogonal side chain protective group was introduced at the position ofthe acylation. The N-terminal of the peptide backbone was thenBoc-protected using Boc2O or alternatively by using a Boc-protectedamino acid in the last coupling. While the peptide was still attached tothe resin, the orthogonal side chain protective group was selectivelycleaved using freshly prepared hydrazine hydrate (2-4%) in NMP for 2×15min. The unprotected lysine side chain was first coupled withFmoc-Glu-OtBu or another spacer amino acid, which was deprotected withpiperidine and acylated with a lipophilic moiety using the peptidecoupling methodology as described above.

Abbreviations employed are as follows:

COMU:1-[(1-(cyano-2-ethoxy-2-oxoethylideneaminooxy)-dimethylamino-morpholinomethylene)]methanaminiumhexaflourophosphate

ivDde: 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)3-methyl-butyl

Dde: 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-ethyl

DCM: dichloromethane

DMF: N,N-dimethylformamide

DIPEA: diisopropylethylamine

EtOH: ethanol

Et₂O: diethyl ether

HATU:N-[(dimethylamino)-1H-1,2,3-triazol[4,5-b]pyridine-1-ylmethylene]-N-methylmethanaminiumhexafluorophosphate N-oxide

MeCN: acetonitrile

NMP: N-methylpyrrolidone

TFA: trifluoroacetic acid

TIS: triisopropylsilane

Cleavage

The resin was washed with EtOH (3×10 ml) and Et₂O (3×10 ml) and dried toconstant weight at room temperature (r.t.). The crude peptide wascleaved from the resin by treatment with TFA/TIS/water (95/2.5/2.5; 40ml, 2 h; r.t.). Most of the TFA was removed at reduced pressure and thecrude peptide was precipitated and washed three times with diethyletherand dried to constant weight at room temperature.

HPLC Purification of the Crude Peptide

The crude peptide was purified to greater than 90% by preparativereverse phase HPLC using a PerSeptive Biosystems VISION Workstationequipped with a C-18 column (5 cm; 10 μm) and a fraction collector andrun at 35 ml/min with a gradient of buffer A (0.1% TFA, aq.) and bufferB (0.1% TFA, 90% MeCN, aq.). Fractions were analyzed by analytical HPLCand MS and relevant fractions were pooled and lyophilized. The finalproduct was characterized by HPLC and MS.

The synthesized compounds are shown in Table 1 (SEQ ID NOs: 1-41).

TABLE 1 Compound No. Sequence  1H-Y-Aib-EGTFISDYSIELDK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS-NH₂ 2H-Y-Aib-EGTFISDYSIELD-K(Hexadecanoyl-isoGlu)-IHQQDFVNWLLAQGPSSGAPPPS-NH₂ 3H-Y-Aib-EGTFISDYSIELEK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS-NH₂ 4 H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂  5H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-  K(Hexadecanoyl-isoGlu)-NH₂  6H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQ-K(Hexadecanoyl-isoGlu)- NH₂  7H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQKG-K(Hexadecanoyl- isoGlu)-NH₂  8H-Y-Aib-EGTFISDYSI ELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNYLLAQGPSSGAPPPS-NH₂  9H-Y-Aib-EGTFISDYSI ELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNWLLAQGPSSGAPPPS-NH₂ 10H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAQGPSSGAPPPS-NH₂ 11H-Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAKEFVNWLLAQGPSSGAPPPS-NH₂ 12H-Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS-NH₂ 13H-Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂ 14H-Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQ-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂ 15H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAGPSSGAPPPS-NH₂ 16H-Y-Aib-EGTFISDYSIELDKIAAQDFVNWLLAGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂ 17H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS-NH₂ 18H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS-NH₂ 19H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS-NH₂ 20H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS-NH₂ 21H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS-NH₂ 22H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS-NH₂ 23H-Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂ 24H-Y-Aib-EGTFISDYSIELDKIAQKEFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂ 25H-Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂ 26H-Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-NH₂ 27H-Y-Aib-EGTFISDYSIELDKIAAQDFVEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂ 28H-Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAQGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂ 29H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQAFVNWLLAGPSSGAPPPS-NH₂ 30H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAAGPSSGAPPPS-NH₂ 31H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAGPSSGAPPPS-NH₂ 32H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS-NH₂ 33H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS-NH₂ 34H-Y-Aib-EGTFISDYSIELD-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-IAQRAFIEWLLAQGPSSGAPPPS-NH₂ 35H-Y-Aib-EGTFISDYS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-ELDKIAQRAFIEWLLAQGPSSGAPPPS-NH₂ 36H-Y-DAla-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS-NH₂ 37H-Y-DAla-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-NH₂ 38H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAQGPSSGAPPPS-NH₂ 39H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAQGPSSGAPPPS-NH₂ 40H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS-NH₂ 41H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS-NH₂Synthesis of Compound No. 9

Solid phase peptide synthesis was performed on a CEM Liberty PeptideSynthesizer using standard Fmoc chemistry. TentaGel S Ram S resin (1.05g; 0.23 mmol/g) was swelled in DMF (10 ml) prior to use and transferredbetween tube and reaction vessel using DCM and DMF.

Coupling

An Fmoc-amino acid in DMF/DCM (2:1; 0.2 M; 5 ml) was added to the resinin a CEM Discover microwave unit together with COMU/DMF (0.5 M; 2 ml)and DIPEA-DMF/DCM (2:1) (2.0 M; 1 ml). The coupling mixture was heatedto 75° C. for 5 min while nitrogen was bubbled through the mixture. Theresin was then washed with DMF (4×10 ml). Fmoc-Tyr(OtBu)-Ser(PsiMe,Me)-OH pseudoproline was used for amino acid number 29 and 30counting from the C-terminal. Lys17 was incorporated as Fmoc-Lys(Dde)-OHfor orthogonal coupling. The first 9 amino acids and amino acid number24 (counting from the C-terminal) was double couple meaning the buildingblock was coupled twice before deprotection. Boc-Tyr(tBu)-OH wasincorporated as the final building block in the N-terminal.

Deprotection

Piperidine/DMF (20%; 10 ml) was added to the resin for initialdeprotection and the mixture was heated by microwaves (30 sec; 40° C.).The reaction vessel was drained and a second portion of piperidine/DMF(20%; 10 ml) was added and heated (75° C.; 3 min.) again. The resin wasthen washed with DMF (6×10 ml).

Side Chain Acylation

While the peptide was still attached to the resin, the orthogonal sidechain protective group was selectively cleaved using freshly preparedhydrazine hydrate (2-4%) in NMP for 2×15 min. The unprotected lysineside chain was first coupled with Fmoc-Glu-OtBu and the two Peg3building blocks using standard coupling and deprotection conditions asexplained above. Lastly the lipophilic moiety was incorporated as a19-carboxy-nonadecanoic acid mono tert butyl ester again using standardcoupling conditions.

Cleavage

The resin was washed with EtOH (3×10 ml) and Et2O (3×10 ml) and dried toconstant weight at room temperature (r.t.). The crude peptide wascleaved from the resin by treatment with TFA/TIS/H₂O (95/2.5/2.5; 60 ml,2 h; r.t.). Most of the TFA was removed at reduced pressure and thecrude peptide was precipitated and washed three times with diethyletherand dried to constant weight at room temperature.

HPLC Purification of the Crude Peptide

The crude peptide was first purified from 30% by preparative reversephase HPLC using a Gilson 331 pump with a Gilson GX281 fractioncollector equipped with a Gemini NX 5μ C-18 110A, 10×250 mm column andrun at 47 ml/min with a gradient of buffer A (0.1% TFA, aq.) and bufferB (0.1% TFA, 90% MeCN, aq.). Fractions were analyzed by analytical HPLCand MS and relevant fractions were pooled and lyophilized. A secondpurification was performed using the same method to obtain the productin 96% purify (53 mg) as characterized by HPLC and MS. Calculatedmonoisotopic mass=5025.54 found 5025.72.

Example 2

Human GIP Receptor (GIP R) Activity Assay

In vitro effects of peptide conjugates of the invention were assessed bymeasuring the induction of cAMP following stimulation of the respectivereceptor by GIP or analogues of these, as outlined in the invention,using the AlphaSceen® cAMP kit from Perkin-Elmer according toinstructions. Briefly, HEK293 cells expressing the human GIP R (stablecell lines generated through transfection of the cDNA for human GIP Rand selection of stable clones) were seeded at 30,000 cells/well in96-well microtiter plates coated with 0.01% poly-L-lysine, and grown for1 day in culture in 200 μl growth medium (DMEM, 10% FCS, Penicillin (100IU/ml), Streptomycin (100 μg/ml)). On the day of analysis, growth mediumwas removed and the cells were washed once with 150 μl Tyrode's buffer(Tyrode's Salts (9.6 g/l), 10 mM HEPES, pH 7.4). Cells were thenincubated in 100 μl Assay buffer (0.1% W/V Alkali-treated Casein and 100μM IBMX in Tyrode's Buffer) containing increasing concentrations ofcontrol and test compounds for 15 min at 37° C. The Assay buffer wasremoved and cells are lysed in 80 μl Lysis buffer (0.1% w/v BSA, 5 mMHEPES, 0.3% v/v Tween-20) per well. From each well 10 μl lysed cells wastransferred to a 384-well plate and mixed with 15 μl bead-mix (1 Unit/15μl anti-cAMP Acceptor Beads, 1 Unit/15 μl Donor Beads, and 1 Unit/15 μlBiotinylated cAMP in Assay Buffer). The plates were mixed and incubatedin the dark for an hour at room temperature before measuring using anEnvision™ plate reader (Perkin-Elmer).

Results were converted into cAMP concentrations using a cAMP standardcurve prepared in KRBH buffer containing 0.1% (v/v) DMSO. The resultingcAMP curves were plotted as absolute cAMP concentrations (nM) over log(test compound concentration) and analyzed using the curve fittingprogram XLfit.

Parameter calculated to describe both the potency as well as theagonistic activity of each test compound on the receptors were:

EC50, a concentration resulting in a half-maximal elevation of cAMPlevels, reflecting the potency of the test compound. The results aresummarized in Table 2.

TABLE 2 EC₅₀ average values of the compounds on the GIP-R compared tocontrol peptide. Compound hGIP-R level of cAMP (nM) hGIP 0.003 1 0.008 20.013 3 0.014 4 0.013 5 0.014 6 0.032 7 0.018 8 0.009 9 0.008 10 0.00711 0.009 12 0.009 13 0.014 14 0.024 15 0.012 16 0.016 17 0.007 18 0.00619 0.006 20 0.007 21 0.007 22 0.005 23 0.010 24 0.008 25 0.032 26 0.01727 0.013 28 0.007 29 0.014 30 0.009 31 0.012 32 0.020 33 0.014 34 0.01135 0.008 36 0.006 37 0.016 38 0.001 39 0.007 40 0.010 41 0.014

Example 3

Activity Assays at Human GIP Receptor (GIP R) and Human GLP-1 Receptor(GLP-1R)

In vitro effects of peptide conjugates were assessed by measuring theinduction of cAMP following stimulation of the respective receptor usingthe AlphaScreen® cAMP kit from Perkin-Elmer according to instructions.Briefly, HEK293 cells expressing the GIP R or the GLP-1R (stable celllines generated through transfection of expression vector containing thecDNA for the receptor in question and selection of stable clones) wereseeded at 30,000 cells/well in 96-well microtiter plates coated with0.01% poly-L-lysine, and grown for 1 day in culture in 200 μl growthmedium (DMEM, 10% FCS, Penicillin (100 IU/ml), Streptomycin (100μg/ml)). On the day of analysis, growth medium was removed and the cellswere washed once with 150 μl Tyrode's buffer (Tyrode's Salts (9.6 g/l),10 mM HEPES, pH 7.4). Cells were then incubated in 100 μl Assay buffer(0.05% W/V Alkali-treated Casein and 100 μM IBMX in Tyrode's Buffer)containing increasing concentrations of control and test compounds for15 min at 37° C. The Assay buffer was removed and cells are lysed in 80μl Lysis buffer (0.1% w/v BSA, 5 mM HEPES, 0.3% v/v Tween-20) per well.From each well 10 μl lysed cells was transferred to a 384-well plate andmixed with 15 μl bead-mix (1 Unit/15 μl anti-cAMP Acceptor Beads, 1Unit/15 μl Donor Beads, and 1 Unit/15 μl Biotinylated cAMP in AssayBuffer). The plates were mixed and incubated in the dark for an hour atroom temperature before measuring using an Envision™ plate reader(Perkin-Elmer).

The cAMP response was normalized relative to a positive and negativecontrol (reference agonist (0.1 nM human GIP or 1 nM Exendin-4 and assaybuffer, respectively) to calculate the EC50 and maximal response fromthe concentration response curve using 4 parameter logistic (4PL)nonlinear regression model for curve fitting.

The EC50s, a concentration resulting in a half-maximal elevation of cAMPlevels, reflecting the potencies of the test agonist compounds aresummarized in Table 2a.

TABLE 2a EC₅₀ average values of the compounds compared to controlpeptides. Compound EC50 hGIP R (nM) EC50 hGLP1 R (nM) Exendin-4 NT 0.004hGIP NT >100 1 0.003 NA 2 0.008 NT 3 0.014 NA 4 0.014 NT 5 0.014 NT 60.014 NT 7 0.032 NA 8 0.019 >10 9 0.009 >3 10 0.008 >10 11 0.008 >3 120.009 >3 13 0.008 >10 14 0.014 >10 15 0.024 >100 16 0.012 >10 170.016 >3 18 0.007 >3 19 0.005 >3 20 0.006 >3 21 0.006 >3 22 0.007 >10 230.005 >10 24 0.010 >10 25 0.008 >100 26 0.032 >10 27 0.017 >10 280.013 >3 29 0.007 >100 30 0.014 >10 31 0.009 >100 32 0.012 >100 330.020 >10 34 0.017 >10 35 0.011 >3 36 0.006 >10 37 0.006 >10 380.017 >100 39 0.001 >100 40 0.007 >10 41 0.010 >10 NT = Not tested, NA =No activity

Example 4

Pharmacokinetics of Selected Compounds in Mice

Method

C57BL/6J mice (males with a body weight of approximately 25 g) weregiven either a single subcutaneous (s.c.) bolus or a single intravenous(i.v.) bolus of each peptide to be tested.

Following s.c. administration of the selected compounds (50, 100 or 200nmol/kg), blood samples were drawn at 8 (eight) timepoints up to 96hours post-dose. Following i.v. administration of the selected compounds(50, 100 or 200 nmol/kg), blood samples were drawn at 8 (eight)timepoints up to 72 hours post-dose. Blood samples were drawn bysublingual bleeding. The dosing vehicle was a phosphate buffercontaining mannitol (pH 7.5).

At each sampling time point, samples from two mice were drawn, i.e. 16mice were included for each compound and each administration route. Themice were euthanized immediately after blood sampling by cervicaldislocation. Plasma samples were analyzed after solid phase extraction(SPE) or protein precipitation followed by liquid chromatography massspectrometry (LC-MS/MS). Mean plasma concentrations were used forcalculation of the pharmacokinetic parameters using thenon-compartmental approach in Phoenix WinNonlin 6.3. Plasma terminalelimination half-life (T½) was determined as In(2)/λz where λz is themagnitude of the slope of the log linear regression of the logconcentration versus time profile during the terminal phase.Bioavailability was determined as AUC_(inf) (s.c.)/AUC_(inf) (i.v.)×100,where AUC_(inf) is the area under the plasma concentration−time curveextrapolated to infinity (AUC_(inf)=AUC_(last)+Clast/λz, where C_(last)is the last observed plasma concentration). T_(max) is the post-dosetime where the maximal plasma concentration was observed. The resultsare summarized in Table 3.

TABLE 3 Terminal elimination half-life (h) and bioavailability in micefollowing s.c. and i.v. administration of selected compounds. T½ (h.)Tmax (h.) Bioavailability Compound i.v. s.c. s.c. s.c. hGIP 0.1 — — — 1016.9 21.1 4  100%*{circumflex over ( )} 12 14.7 16.8 8  77%{circumflexover ( )} 15 19.2 16.7 8 87% 16 23.3 23.6 8 81% 13 14.4 13.7 8 75% 1619.2 16.7 8 88% 17 16.3 19.9 8 56% 18 17.6 15.1 4 78% 21 24.8 21.0 8 67%33 21.7 18.7 8 78% 35 14.5 14.5 4 73% 41 17.6 16.5 8 70% *Thebioavailability was capped to 100% {circumflex over ( )}In a repeatedtest the bioavalability of Compound 10 was 77% and the bioavailabilityof Compound 12 was 98%.

Example 5

OGTT (Oral Glucose Tolerance Test) in Normal Mice.

Male C57BL/6J mice (Charles River, Germany) were maintained on normalchow (Altromin 1324, Brogaarden A/S, Gentofte, Denmark) and domesticquality water with added citric acid to pH≠3.6. The animals were housedin groups of n=3 in a light-, temperature-, and humidity-controlled room(12:12 h light-dark cycle, with lights on at 06.00-18.00 hr; 21±1° C.;50-80% relative humidity). Mice, 10-12 weeks old, were fasted 5 hoursbefore the OGTT. GIP receptor agonists (3-300 nmol/kg), the GLP-1analogue liraglutide (10 nmol/kg) and vehicle were administered (5mL/kg) subcutaneously (s.c.) 4 hours before the oral gavage of glucose(t=0 min; 2 g/kg; 5 mL/kg). Tail vein blood was sampled at time t=0(before glucose administration), 15, 30, 60, and 120 min formeasurements of blood glucose. Results (blood glucose levels and areaunder the blood glucose curves (AUC). Data are means±SEM; n=6) from 2experiments are shown in FIGS. 1 (A-D) and 2 (A-E).

Statistical analyses were performed using Graph Pad Prism version 5. Theblood glucose AUCs were compared using one-way ANOVA followed byDunnett's Multiple Comparison

Tests vs. vehicle group. Differences were considered statisticallysignificant at p<0.05. Statistical differences vs vehicle: *p<0.05,**p<0.01, ***p<0.001.

Example 6

Sub-Chronic Effects of Co-Treatment of GIP Receptor Agonist and GLP-1Receptor Agonist on Body Weight in Diet-Induced Obese (DIO) C57BL/6JMice

Male C57BL/6J (JAX) mice (Charles River, UK) fed high-fat diet (45% oftotal energy from fat, D12451 Research Diet Inc.) for approximately 4months were used. The animals were housed in groups of n=3 in a light-,temperature-, and humidity-controlled room (12:12 h light-dark cycle,with lights on at 07.00-19.00 hr; 21±2° C.; 55±20% relative humidity).Mice were single-housed two weeks prior to start of the mock phase. Allmice were mock-treated (once daily s.c. injection of vehicle) for a weekto acclimatize the animals to handling and injections. Subsequently, themice were stratified according to body weight into treatment groups(n=8-9). The average starting body weight was 39-40 grams. Animals werethereafter treated once daily with two separate s.c. injections (3 mL/kgof each injection) from day 1 to day 22. The first injection was withvehicle 1 (25 mM phosphate, 125 mM sodium chloride buffer, pH 7.4) orGLP-1 analogue liraglutide (20 nmol/kg). The second injection was withvehicle 2 (25 mM phosphate, 205 mM D-Mannitol, pH 7.5) or GIP agonist (3and 30 nmol/kg). The GIP agonist was only dosed every third day of thestudy (starting on day 1). On other days, GIP agonist was replaced withvehicle 2. The daily injections were given in the morning (at9.00-10.00). Body weight was determined daily throughout the study.Changes in body weight during the study are shown in FIG. 3 (deltaΔ=body weight at each study day−body weight at day 1. Data aremeans±SEM).

Statistical analyses were performed using Graph Pad Prism version 5. Thechange in body weight of liraglutide-treated mice was compared with miceco-administered liraglutide and GIP agonist by two-way ANOVA followed byBonferroni posttests. P<0.05 was considered statistically significant.The change in body weight of vehicle-treated control mice was comparedwith compound-treated mice by two-way ANOVA followed by Bonferroniposttests; ***p<0.001 vs. vehicle. Statistical differences vs vehicleare shown for day 22 in FIG. 3.

Example 7

Sub-Chronic Effects of Co-Treatment of GIP Receptor Agonists and GLP-1Receptor Agonist on Body Weight in Diet-Induced Obese (DIO) C57BL/6JMice

Male C57BL/6J mice (Charles River, Germany) fed high-fat diet (60% oftotal energy from fat, DIO Rodent Purified 58Y1-58126 from TestDiet) forapproximately 5 months were used. The animals were housed in groups ofn=3 in a light-, temperature-, and humidity-controlled room (12:12 hlight-dark cycle, with lights on at 06.00-18.00 hr; 21±1° C.; 65±15%relative humidity). All mice were mock-treated (once daily s.c.injection of vehicle) for a week to acclimatize the animals to handlingand injections. Subsequently, the mice were stratified according to bodyweight into treatment groups (n=9). The average starting body weight was40-41 grams. Animals were thereafter treated once daily with twoseparate s.c. injections (5 mL/kg of each injection) from day 0 to day27. The first injection was with vehicle 1 (25 mM phosphate, 125 mMsodium chloride buffer, pH 7.4) or GLP-1 analogue liraglutide (20nmol/kg). The second injection was with vehicle 2 (25 mM phosphate, 205mM D-Mannitol, pH 7.5) or GIP agonist (30 and/or 300 nmol/kg). The GIPagonist was only dosed every third day of the study (starting on day 0).On other days, GIP agonist was replaced with vehicle 2. The dailyinjections were given in the morning (at 9.00-10.00). Body weight wasdetermined daily throughout the study. Changes in body weight during thestudy (delta Δ body weight=body weight at each study day−body weight atday 0. Data are means±SEM) are shown in FIG. 4 A (Compound 10 and 12), B(Compound 17), C (Compound 18), D (compound 35) and E (Compound 41).

Statistical analyses were performed for using Graph Pad Prism version 5.The change in body weight of liraglutide-treated mice was compared withmice co-administered liraglutide and GIP agonist by two-way ANOVAfollowed by Bonferroni posttests. P<0.05 was considered statisticallysignificant (illustrated with lines below the body weight curves). Thechange in body weight of vehicle-treated control mice was compared withcompound-treated mice by two-way ANOVA followed by Bonferroni posttests;***p<0.001 vs. vehicle. Statistical differences vs vehicle are shown forday 27 (FIG. 4 A-E).

The invention claimed is:
 1. A GIP analogue represented by the generalFormula I:R¹-Tyr-X2-Glu-Gly-Thr-Phe-Ile-Ser-Asp-X10-X11-X12-Glu-Leu-X15-X16-X17-X18-X19-X20-X21-Phe-X23-X24-X25-Leu-X27-X28-X29-Y1-Y2-R²  (I)(SEQ ID NO: 42) wherein R¹ is H—, Ac or pGlu; X2 is Aib or D-Ala; X10 isTyr; X11 is Ser; X12 is Ψ or Ile; X15 is Asp or Glu; X16 is Lys or Ψ;X17 is Ile or Ψ; X18 is His or Ala; X19 is Gln or Ala; X20 is Gln, Lys,or Arg; X21 is Ala, Asp or Glu; X23 is Val or Ile; X24 is Asn or Glu;X25 is Tyr or Trp; X27 is Leu; X28 is Ala; X29 is Ala, Gln, Thr, Ser orLys or is absent; Y1 is Lys-Gly, Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser(SEQ ID NO: 43), Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 44),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser (SEQ ID NO: 45),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 46), or absent; Y2 is Ψ oris absent; R² is —NH₂ or —OH; wherein Ψ is a Lys residue and wherein theside chain of said Lys residue is conjugated to a lipophilicsubstituent; wherein the GIP analogue contains one and only one residueΨ; and wherein the GIP analogue has agonist activity at the GIPreceptor; or a pharmaceutically acceptable salt thereof.
 2. A GIPanalogue according to claim 1 comprising one of the following residuesor combinations of residues: Aib2, Asp15, Lys20; Aib2, Asp15, Arg20;Aib2, Asp15, Arg20, Ile23; Aib2, Ile12, Asp15, Arg20, Ile23, Glu24;Ile12, Asp15, Ile23; Ile12, Asp15, Ile23 Glu24; Ile12, Asp15, Ala21,Ile23; Aib2, Ala21, Ile23, Glu24; Aib2, Asp15, Ile23; Aib2, Asp15,Arg20, Ile23, Gln29; DAla2, Asp15, Ile23; Aib2, Asp15, Ile17, Lys20,Ala28; Asp15, Ile23, Glu24.
 3. A GIP analogue according to claim 1represented by the general Formula III:R¹-Tyr-Aib-Glu-Gly-Thr-Phe-Ile-Ser-Asp-Tyr-Ser-Ile-Glu-Leu-X15-X16-X17-X18-X19-X20-X21-Phe-Val-X24-X25-Leu-Leu-Ala-X29-Y1-Y2-R²  (III)(SEQ ID NO: 50) wherein R¹ is H—, Ac or pGlu; X15 is Asp or Glu; X16 isLys or Ψ; X17 is Ile or Ψ; X18 is His or Ala X19 is Gln or Ala; X20 isGln, Lys or Arg; X21 is Ala, Asp or Glu; X24 is Asn or Glu; X25 is Tyror Trp X29 is Gln or is absent; Y1 is Lys-Gly,Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser (SEQ ID NO: 43),Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 44),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser (SEQ ID NO: 45),Pro-Ser-Ser-Gly-Ala-Pro-Pro-Ser (SEQ ID NO: 46), or absent; Y2 is Ψ oris absent; R² is —NH₂ or —OH; wherein Ψ is a Lys residue and wherein theside chain of said Lys residue is conjugated to a lipophilicsubstituent; and wherein the GIP analogue contains one and only oneresidue Ψ; and wherein the GIP analogue has agonist activity at the GIPreceptor; or a pharmaceutically acceptable salt thereof.
 4. A GIPanalogue according to claim 3 comprising one of the following residuesor combinations of residues: Asp15, Lys20; Asp15, Arg20; Asp15, Arg20,Glu24; Asp15, Lys16; Asp15, Lys16, Glu24; Asp15, Ψ16, Ala21; Ala21,Glu24; Asp15, Arg20, Gln29; Asp15, Arg20, Gly29; Asp15, Ile17, Arg20,Gly29; Asp15, Ile17, Lys20, Gly29; Asp15, Ala28; Asp15, Ile17, Lys20,Ala28; Asp15, Ile23, Glu24; Asp15, Ψ17, Lys20; Asp15, Ψ17, Arg20; Asp15, Ψ17, Arg20 Asp15, Ψ17, Arg20, Glu24; Asp15, Lys 16, Ψ17; Asp15, Lys16, Ψ17, Glu24; Asp15 , Ψ17, Ala21; Ala21, Ψ17, Glu24; Asp15, Asp15 ,Ψ17, Arg20, Gln29; Asp15 , Ψ17, Arg20, Gly29; Asp15, Ile17, Arg20,Gly29; Asp15, Ile17, Lys20, Gly29; Asp15; Ψ17; Asp15, Ψ17, Ala28; Asp15,Ile17, Lys20, Ala28; Asp15 , Ψ17, Ile23, Glu24.
 5. A GIP analogueaccording to claim 1, wherein Ψ is a Lys in which the side chain isconjugated to a substituent having the formula —Z¹ or —Z²—Z¹.
 6. A GIPanalogue according to claim 5, wherein —Z¹ is a fatty chain having at aterminus a connection —X— to Ψ or to Z²; wherein —X— is a bond, —CO—,—SO—, or —SO₂—; and, optionally, Z¹ has a polar group at the end of thechain distal from connection —X—; said polar group comprising acarboxylic acid or a carboxylic acid bioisostere, a phosphonic acid, ora sulfonic acid group.
 7. A GIP analogue according to claim 6 wherein Z¹is a group of formula: A-B-Alk-X— wherein A is hydrogen or a carboxylicacid, a carboxylic acid bioisostere, a phosphonic acid, or a sulfonicacid group; B is a bond or a linker; X is a bond, acyl (—CO—), sulfinyl(—SO—), or sulfonyl (—SO₂—); and Alk is a fatty chain that may beoptionally substituted with one or more substituents.
 8. A GIP analogueaccording to claim 7, wherein Z¹ is: A-B—C₁₆₋₂₀ alkylene-(CO)— wherein Ais H or —COOH and B is 17-carboxy-heptadecanoyl HOOC—(CH₂)₁₆—(CO)—;19-carboxy-nonadecanoyl HOOC—(CH₂)₁₈—(CO)—; OctadecanoylH₃C—(CH₂)₁₆—(CO)—; or Eicosanoyl H₃C—(CH₂)₁₈—(CO)—.
 9. A GIP analogueaccording to claim 5, wherein Z² is a spacer bound at one terminus by Y,which is a nitrogen, oxygen or sulfur atom, and at the other terminus byX, which is a bond or an acyl (—CO—), sulfinyl (—SO—), sulfonyl (—SO₂—)or absent.
 10. A GIP analogue according to claim 5, wherein —Z¹-Z² is:


11. A GIP analogue according to claim 1 having the sequence(SEQ ID NO: 51) Y-Aib-EGTFISDYSIELDKΨHQQDFVNWLLAQGPSSGAPPPS;(SEQ ID NO: 52) Y-Aib-EGTFISDYSIELDΨIHQQDFVNWLLAQGPSSGAPPPS;(SEQ ID NO: 53) Y-Aib-EGTFISDYSIELEKΨHQQDFVNWLLAQGPSSGAPPPS;(SEQ ID NO: 54) Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPSΨ;(SEQ ID NO: 55) Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQΨ; (SEQ ID NO: 56)Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQKGΨ; (SEQ ID NO: 57)Y-Aib-EGTFISDYSIELDKΨHQQDFVNYLLAQGPSSGAPPPS; (SEQ ID NO: 58)Y-Aib-EGTFISDYSIELDKΨHQQDFVNWLLAQGPSSGAPPPS; (SEQ ID NO: 69)Y-Aib-EGTFISDYSIELDKΨAAQDFVNWLLAQGPSSGAPPPS; (SEQ ID NO: 60)Y-Aib-EGTFISDYSIELEKΨAAKEFVNWLLAQGPSSGAPPPS; (SEQ ID NO: 61)Y-Aib-EGTFISDYSIELEKΨAQRAFVEWLLAQGPSSGAPPPS; (SEQ ID NO: 62)Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQGPSSGAPPPSΨ; (SEQ ID NO: 63)Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQΨ; (SEQ ID NO: 64)Y-Aib-EGTFISDYSIELDKΨAAQDFVNWLLAGPSSGAPPPS; (SEQ ID NO: 65)Y-Aib-EGTFISDYSIELDKIAAQDFVNWLLAGPSSGAPPPSΨ; (SEQ ID NO: 66)Y-Aib-EGTFISDYSIELDKKΨAQRAFVEWLLAQGPSSGAPPPS; (SEQ ID NO: 67)Y-Aib-EGTFISDYSIELDKKΨAQRAFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 68)Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 69)Y-Aib-EGTFISDYSIELDKIAQKEFIEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 70)Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 71)Y-Aib-EGTFISDYSIELDKIAAQDFVEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 72)Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAQGPSSGAPPPSKΨ; (SEQ ID NO: 73)Y-Aib-EGTFISDYSIELDKKΨAAQAFVNWLLAGPSSGAPPPS; (SEQ ID NO: 74)Y-Aib-EGTFISDYSIELDKKΨAAQDFVNWLLAAGPSSGAPPPS, (SEQ ID NO: 75)Y-Aib-EGTFISDYSIELDKKΨAAQDFINWLLAGPSSGAPPPS, (SEQ ID NO: 76)Y-Aib-EGTFISDYSIELDKKΨAAQDFIEWLLAGPSSGAPPPS, (SEQ ID NO: 77)Y-Aib-EGTFISDYSIELDKKΨAAQDFIEWLLAGPSSGAPPPS, (SEQ ID NO: 78)Y-Aib-EGTFISDYSIELDKΨIAQRAFIEWLLAQGPSSGAPPPS, (SEQ ID NO: 79)Y-Aib-EGTFISDYSKΨELDKIAQRAFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 80)Y-DAla-EGTFISDYSIELDKKΨAQRAFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 81)Y-DAla-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPSKΨ; (SEQ ID NO: 82)Y-Aib-EGTFISDYSIELDKKΨAAQDFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 83)Y-Aib-EGTFISDYSIELDKKΨAAQDFINWLLAQGPSSGAPPPS,  or (SEQ ID NO: 84)Y-Aib-EGTFISDYSIELDKKΨAAQAFIEWLLAQGPSSGAPPPS.


12. A GIP analogue according to claim 1 having the sequenceY-Aib-EGTFISDYSIELDK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 1);Y-Aib-EGTFISDYSIELD-K(Hexadecanoyl-isoGlu)-IHQQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 2);Y-Aib-EGTFISDYSIELEK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 3);Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 4);Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-K(Hexadecanoyl-isoGlu) (SEQ ID NO: 5);Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQ-K(Hexadecanoyl-isoGlu) (SEQ ID NO: 6);Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQKG-K(Hexadecanoyl-isoGlu) (SEQ ID NO: 7);Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNYLLAQGPSSGAPPPS (SEQ ID NO: 8);Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 9);Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAQGPSSGAPPPS (SEQ ID NO: 10);Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAKEFVNWLLAQGPSSGAPPPS (SEQ ID NO: 11);Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS (SEQ ID NO: 12);Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 13);Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQ-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 14);H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAGPSSGAPPPS (SEQ ID NO: 15);Y-Aib-EGTFISDYSIELDKIAAQDFVNWLLAGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 16);Y-A1b-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS (SEQ ID NO: 17);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 18);Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 19);Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS (SEQ ID NO: 20);Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS (SEQ ID NO: 21);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS (SEQ ID NO: 22);Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 23);Y-Aib-EGTFISDYSIELDKIAQKEFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 24);Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 25);Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3) (SEQ ID NO: 26);Y-Aib-EGTFISDYSIELDKIAAQDFVEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 27);Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAQGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3) (SEQ ID NO: 28);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyn-isoGlu-Peg3-Peg3)-AAQAFVNWLLAGPSSGAPPPS (SEQ ID NO: 29);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAAGPSSGAPPPS (SEQ ID NO: 30);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAGPSSGAPPPS (SEQ ID NO: 31);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS (SEQ ID NO: 32);Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS (SEQ ID NO: 33);Y-Aib-EGTFISDYSIELD-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-IAQRAFIEWLLAQGPSSGAPPPS; (SEQ ID NO: 34)Y-Aib-EGTFISDYS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-ELDKIAQRAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 35);Y-DAla-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 36);Y-DAla-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3) (SEQ ID NO: 37);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAQGPSSGAPPPS (SEQ ID NO: 38);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAQGPSSGAPPPS (SEQ ID NO: 39);Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 40); orY-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS (SEQ ID NO: 41).


13. A GIP analogue according to claim 1 which is: (SEQ ID NO: 1)H-Y-Aib-EGTFISDYSIELDK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS-NH₂;(SEQ ID NO: 2)H-Y-Aib-EGTFISDYSIELD-K(Hexadecanoyl-isoGlu)-IHQQDFVNWLLAQGPSSGAPPPS-NH₂;(SEQ ID NO: 3)H-Y-Aib-EGTFISDYSIELEK-K(Hexadecanoyl-isoGlu)-HQQDFVNWLLAQGPSSGAPPPS-NH₂;(SEQ ID NO: 4)H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 5)H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQGPSSGAPPPS-K(Hexadecanoyl-isoGlu)-NH₂;(SEQ ID NO: 6)H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQ-K(Hexadecanoyl-isoGlu)-NH₂;(SEQ ID NO: 7)H-Y-Aib-EGTFISDYSIELDKIHQQDFVNWLLAQKG-K(Hexadecanoyl-isoGlu)-NH₂;(SEQ ID NO: 8)H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNYLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 9)H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-HQQDFVNWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 10)H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 11)H-Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAKEFVNWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 12)H-Y-Aib-EGTFISDYSIELEK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 13)H-Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 14)H-Y-Aib-EGTFISDYSIELEKIAQRAFVEWLLAQ-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 15)H-Y-Aib-EGTFISDYSIELDK-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 16)H-Y-Aib-EGTFISDYSIELDKIAAQDFVNWLLAGPSSGAPPPS-K([19-carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 17)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 18)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 19)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAF1EWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 20)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQRAFVEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 21)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS-NH₂; (SEQ ID NO: 22)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQKEFVEWLLAAGPSSGAPPPS-NH₂; (SEQ ID NO: 23)H-Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 24)H-Y-Aib-EGTFISDYSIELDKIAQKEFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 25)H-Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 26)H-Y-Aib-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyI)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-NH₂; (SEQ ID NO: 27)H-Y-Aib-EGTFISDYSIELDKIAAQDFVEWLLAGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 28)H-Y-Aib-EGTFISDYSIELDKIAQRAFIEWLLAQGPSSGAPPPS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-NH₂; (SEQ ID NO: 29)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQAFVNWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 30)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFVNWLLAAGPSSGAPPPS-NH₂; (SEQ ID NO: 31)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 32)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 33)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQDFIEWLLAGPSSGAPPPS-NH₂; (SEQ ID NO: 34)H-Y-Aib-EGTFISDYSIELD-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-IAQRAFIEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 35)H-Y-Aib-EGTFISDYS-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-ELDKIAQRAFIEWLLAQGPSSGAPPPS-NH_(2;;); (SEQ ID NO: 36)H-Y-DAla-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AQRAFIEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 37)H-Y-DAla-EGTFISDYSIELDKIAAQDFIEWLLAGPSSGAPPPS-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-NH₂; (SEQ ID NO: 38)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFIEWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 39)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQDFINWLLAQGPSSGAPPPS-NH₂; (SEQ ID NO: 40)H-Y-Aib-EGTFISDYSIELDK-K([19-Carboxy-nonadecanoyl]-isoGlu-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS-NH₂;  and or (SEQ ID NO: 41)H-Y-Aib-EGTFISDYSIELDK-K((19-Carboxy-nonadecanoyl)-[(Piperazine-1-yl)-acetyl]-Peg3-Peg3)-AAQAFIEWLLAQGPSSGAPPPS-NH2;.


14. A pharmaceutical composition comprising a GIP analogue according toclaim 1, or a pharmaceutically acceptable salt thereof, in admixturewith a carrier.
 15. A method of treating a metabolic disorder in anindividual in need thereof, comprising administering to said individuala GIP analogue according to claim 1, or a pharmaceutically acceptablesalt thereof.
 16. The method according to claim 15 wherein the metabolicdisorder is diabetes, a diabetes related disorder, obesity, or anobesity related disorder.
 17. The method according to claim 16 whereinthe diabetes related disorder is selected from the group consisting ofinsulin resistance, glucose intolerance, increased fasting glucose,hypoglycemia (for example induced by insulin treatment), pre-diabetes,type 1 diabetes, type 2 diabetes, gestational diabetes hypertension anddyslipidemia.
 18. The method according to claim 16 wherein the obesityrelated disorder is selected from the group consisting of obesity linkedinflammation, obesity linked gallbladder disease, obesity induced sleepapnea, atherogenic dyslipidemia, blood fat disorders, elevated bloodpressure, hypertension, a prothrombotic state, a proinflammatory state,and combinations thereof.